Lab38 Acred. de Labs. Microbiológicos PDF

LAB 38 Accreditation for microbiological laboratories
Edition 1 September 2001 Page A (+ pp 1 - 28)

Section Page
Contents
Accreditation for
Microbiological Laboratories
[UKAS Publication ref: LAB 38]
Preface B
1 Introduction 4
2 Scope of Accreditation (ISO/IEC 17025, section 1, para. 4.1.2, and UKAS Agreement) 5
3 Staff (ISO/IEC 17025, section 5.2) 5
4 Environment (ISO/IEC 17025, section 5.5) 6
5 Equipment maintenance, calibration and performance verification
(ISO/IEC 17025, section 5.5) 9
6 Reagents and culture media (ISO/IEC 17025, sections 4.6 and 5.5) 8
7 Test methods and procedures (ISO/IEC 17025, section 5.4.1-5.4.4) 14
8 Validation of methods and verification of performance (ISO/IEC 17025, section 5.4.5) 14
9 Quality assurance of results/quality control (ISO/IEC 17025, section 5.9) 15
10 Laboratory audit and review (ISO/IEC 17025, sections 4.10, 4.13 and 4.14) 17
11 Sample handling and identification (ISO/IEC 17025, sections 5.7 and 5.8) 18
12 Disposal of contaminated waste (ISO/IEC 17025, section 5.8.1) 19
13 Uncertainty of measurement (ISO/IEC 17025, section 5.4.6) 19
14 Use of computers (ISO/IEC 17025, section 5.4.7) 19
Appendix A Glossary of terms 20
Appendix B Bibliography 22
Appendix C References 22
Appendix D Use of reference cultures (ISO/IEC 17025, section 5.6.2.2.2 and 5.6.3.2) 23
Appendix E Guidance on calibration and calibration checks
(ISO/IEC 17025, sections 5.5 and 5.6) 24
Appendix F Guidance on commissioning and verification of performance
(ISO/IEC 17025, section (ISO/IEC 17025, section 5.5.2 and 5.5.5) 25
Appendix G Guidance on maintenance of equipment (ISO/IEC 17025, section 5.5) 27
LAB 38 Accreditation for microbiological laboratories
Page B (+ pp 1 - 28) Edition 1 September 2001
LAB 38 Edition 1 September 2001
United Kingdom Accreditation Service
21 47 High Street
Feltham
Middlesex TW13 4UN
UK
Tel: 020 8917 84000
Fax: 020 8917 8500
website: www.ukas.com
United Kingdom Accreditation Service 2001
About the United Kingdom Accreditation Service
The United Kingdom Accreditation Service (UKAS) is recognised by the
UK Government as the national body responsible for assessing and accrediting the
competence of organisations in the fields of calibration, testing, inspection and
certification of systems, products and personnel.
Preface
ISO/IEC 17025 contains the requirements that testing and calibration laboratories
have to meet if they wish to demonstrate that they operate to a quality system, are
technically competent and are able to generate technically valid results. It replaces
ISO/IEC Guide 25 and EN 45001, and is the standard which UKAS will now use
in place of UKAS publication M10 to assess a laboratory's competence for the
purposes of accreditation.
The publication that follows is EAL Guidance Document No. 18 (current EA
reference number EA-4/10) which was produced by the joint EAL/EURACHEM
Chemistry Working Group to which UKAS (as NAMAS) was an active
contributor. The publication makes reference to EN 45001, EN 45002 and ISO/IEC
Guide 25 throughout the text. Although these standards are been replaced by
ISO/IEC 17025, the document contains continued guidance on implementation of
accreditation in chemical laboratories.
The publication is also available as a free download from the website to the
European cooperation for Accreditation (EA) at www.european-accreditation.org
(publication reference EA-4/10).
UKAS has continued to adopt the publication, now as LAB 38, Edition 1. LAB 38
supersedes UKAS (NAMAS) publication NIS 31, which has been withdrawn.
To assist in using this document for further guidance, cross- referencing of ISO/
IEC 17025 requirements to the appropriate sections of the Eurachem Guidance
Document No. 1/WELAC Guidance document No. WGD 2 (Edition 1) are given
on page A.
PAGE 1 OF 28 EDI TI ON 1 MAY 1996
EAL-G18 ACCREDI TATI ON FOR MI CROBI OLOGI CAL LABORATORI ES
EAL-G18
Publication Reference
Accredi tati on for Laboratori es
Performi ng Mi crobi ol ogi cal
Testi ng
PURPOSE
Thi s document has been produced by a joi nt EAL/EURACHEM Worki ng Group. I t suppl ements
EN 45001 and I SO/I EC Gui de 25 and pr ovi des speci fi c gui dance on the accr edi tati on of
l aboratori es performi ng mi crobi ol ogi cal testi ng, for both assessors and l aboratori es prepari ng
for accredi tati on. EN 45001 and I SO/I EC Gui de 25 remai n the authori tati ve documents and,
i n cases of di spute, the i ndi vi dual accredi tati on bodi es wi l l adjudi cate on unresol ved matters.
The gui dance gi ven i n thi s document may al so be of use to those worki ng towards certi fi cati on
to the I SO 9000 (EN 29000) seri es of standards.
-KH=?DA
EAL
European cooperation for
Accreditation of
Laboratories
EDI TI ON 1 MAY 1996 PAGE 2 OF 28
Calib National member Testing National
member
Austria BMwA BMwA
Tel: +431 711 02 233 Tel: +431 711 02 248
Fax: +431 714 35 82 Fax: +431 714 35 82
Belgium BKO/OBE BELTEST
Tel: +32 2 206 47 35 Tel: +32 2 206 46 80
Fax: +32 2 206 57 45 Fax: +32 2 206 57 42
Denmark DANAK DANAK
Tel: +45 35 86 86 86 Tel: +45 35 86 82 80
Fax: +45 35 86 86 87 Fax: +45 35 86 86 87
Finland FINAS FINAS
Tel: +358 0 616 71 Tel: +358 0 616 71
Fax: +358 0 616 7341 Fax: +358 0 616 7467
France COFRAC COFRAC
Tel: +33 1 44 68 82 33 Tel: +33 1 44 68 82 40
Fax: +33 1 44 68 82 22 Fax: +33 1 44 68 82 23
Germany DKD DAR
Tel: +49 531 592 8320 Tel: +49 30 8104 1710
Fax: +49 531 592 9292 Fax: +49 30 8104 1717
Greece Ministry of Commerce ELOT
Tel: +30 1 38 14 168 Tel: +30 1 201 5025
Fax: +30 1 38 42 642 Fax: +30 1 202 0776
Iceland ISAC ISAC
Tel: +354 1 681 122 Tel: +354 1 681 122
Fax: +354 1 685 998 Fax: +354 1 685 998
Ireland NAB NAB
Tel: +353 1 607 3003 Tel: +353 1 607 3003
Calib National member Testing National
member
Fax: +353 1 607 3109 Fax: +353 1 607 3109
Italy SIT SINAL
Tel: +39 11 348 8933 Tel: +39 6 487 1176
Fax: +39 11 348 6384 Fax: +39 6 481 4563
Netherlands RvA RvA
Tel: +3130 239 4500 Tel: +3130 239 4500
Fax: +3130 239 4539 Fax: +3130 239 4539
Norway NA NA
Tel: +47 22 20 0226 Tel: +47 22 20 0226
Fax: +47 22 20 7772 Fax: +47 22 20 7772
Portugal IPQ IPQ
Tel: +351 1 294 8201 Tel: +351 1 294 8201
Fax: +351 1 294 8202 Fax: +351 1 294 8202
Spain ENAC ENAC
Tel: +34 1 457 32 89 Tel: +34 1 457 32 89
Fax: +34 1 458 62 80 Fax: +34 1 458 62 80
Sweden SWEDAC SWEDAC
Tel: +46 33 17 77 30 Tel: +46 84 02 00 71
Fax: +46 33 10 13 92 Fax: +46 87 91 89 29
Switzerland SAS SAS
Tel: +41 31 963 3381 Tel: +41 31 963 3412
Fax: +41 31 963 3210 Fax: +41 31 963 3210
United Kingdom UKAS UKAS
Tel: +44 181 943 7068 Tel: +44 181 943 6266
Fax: +44 181 943 7134 Fax: +44 181 943 7134
Authorship
This publication has been written jointly by EAL and EURACHEM
Official language
The text may be translated into other languages as required. The English language version remains
the definitive version.
Copyright
The copyright of this text is held by EAL and EURACHEM. The text may not be copied for resale.
Guidance Publications
This document represents a consensus of EAL member opinion and preferred practice on how the
relevant clauses of the accreditation standards might be applied in the context of the subject matter
of this document. The approaches taken are not mandatory and are for the guidance of accreditation
bodies and their client laboratories. Nevertheless, the document has been produced as a means of
promoting a consistent approach to laboratory accreditation amongst EAL member bodies,
particularly those participating in the EAL Multilateral Agreement.
Further information
For further information about this publication, contact your National member of EAL, whose telephone
and facsimile numbers are given below:
Section Page
Contents
1 I ntroducti on 4
2 Scope of accredi tati on 5
3 Staff 5
4 Envi ronment 6
5 Equi pment mai ntenance, cal i brati on and performance veri fi cati on 9
6 Reagents and cul ture medi a 12
7 Test methods and procedures 14
8 Val i dati on of methods and veri fi cati on of performance 14
9 Qual i ty assurance of resul ts/qual i ty control 15
10 Laboratory audi t and revi ew 17
11 Sampl e handl i ng and i denti fi cati on 18
12 Di sposal of contami nated waste 19
13 Uncertai nty of measurement 19
14 Use of computers 19
Appendi x A Gl ossary of terms 20
Appendi x B Bi bl i ography 22
Appendi x C References 22
Appendi x D Use of reference cul tures 23
Appendi x E Gui dance on cal i brati on and cal i brati on checks 24
Appendi x F Gui dance on commi ssi oni ng and veri fi cati on of performance 25
Appendi x G Gui dance on mai ntenance of equi pment 27
1 Introduction and scope of document
1.1 The gener al r equi r ements for accr edi tati on ar e l ai d down i n the Eur opean
Standar d General cri teri a for the operati on of testi ng l aboratori es
(EN 45001:1989) and General requirements for the competence of calibration and
testing laboratories (I SO/I EC Gui de 25, 3rd Ed., 1990), hereafter referred to as
EN 45001 and I SO Gui de 25 respecti vel y. Al l of these requi rements must be
met by l aboratori es seeki ng accredi tati on.
1.2 Thi s document onl y suppl ements EN 45001 and I SO Gui de 25 by pr ovi di ng
speci fi c gui dance for both assessor s and for l abor ator i es car r yi ng out
mi cr obi ol ogi cal testi ng. I t gi ves detai l ed gui dance on the i nter pr etati on of
EN 45001 and I SO Gui de 25 for those undertaki ng the exami nati on of materi al s,
products and substances. The gui dance i s appl i cabl e to the performance of al l
objecti ve measurements, whether routi ne, non-routi ne, or as part of research
and devel opment. EN 45001 and I SO Gui de 25 r emai n the author i tati ve
documents and, i n cases of di spute, accr edi tati on bodi es wi l l adjudi cate on
unresol ved matters. The gui dance gi ven i n thi s document may al so be of use to
those worki ng towards regi strati on under other qual i ty standards such as GLP,
GMP, GCP and certi fi cati on to I SO 9000.
1.3 Mi crobi ol ogi cal testi ng i s taken to i ncl ude steri l i ty testi ng, detecti on, i sol ati on,
enumer ati on and i denti fi cati on of mi cr o-or gani sms (vi r uses, bacter i a,
mycopl asms, fungi and protozoa) and thei r metabol i tes i n di fferent materi al s
and products, or any ki nd of assay usi ng mi cro-organi sms as part of a detecti on
system as wel l as the use of mi cro-organi sms for ecol ogi cal testi ng. I t fol l ows
that some of the gui dance i n thi s document, eg on steri l i ty of the l aboratory
envi ronment, wi l l need to be i nterpreted accordi ngl y. Thi s document can al so
pr ovi de gui dance to l abor ator i es usi ng techni ques i n ar eas r el ated to
mi crobi ol ogy, such as bi ochemi stry, mol ecul ar bi ol ogy and cel l cul ture. However,
there may al so be addi ti onal requi rements for l aboratori es i n areas not covered
by thi s document
1.4 Thi s document i s concerned wi th the qual i ty of test resul ts and i s not speci fi cal l y
concerned wi th heal th and safety matters. However, l aboratory practi ces shoul d
conform to nati onal heal th and safety regul ati ons. I t i s i mportant to note that
i n some cases heal th and safety i ssues may have an effect on qual i ty and the
l aboratory wi l l be requi red to take thi s i nto account.
1.5 Defi ni ti ons of the terms used are gi ven i n Appendi x A.
2 Scope of accreditation
(EN45002, paragraphs 4 and 12)
2.1 The scope of accredi tati on of a l aboratory i s the formal statement of the range
of acti vi ti es for whi ch the l aboratory has been accredi ted; the scope i s recorded
on an accr edi tati on schedul e whi ch i s i ssued together wi th the l abor ator ys
accredi tati on certi fi cate. The l aboratorys scope shoul d be defi ned as preci sel y
as possi bl e so that al l parti es concerned know accuratel y and unambi guousl y
the r ange of tests and/or anal yses cover ed by that par ti cul ar l abor ator ys
accredi tati on.
3 Staff
EN45001, paragraph 5.2
ISOGuide 25, paragraph 6
3.1 The l aboratory management shoul d defi ne the mi ni mum l evel s of qual i fi cati on
and experi ence necessary for al l posts wi thi n the l aboratory.
3.2 Mi crobi ol ogi cal testi ng must be ei ther performed or supervi sed by an experi enced
person, qual i fi ed to degree l evel i n mi crobi ol ogy (or equi val ent). Al ternati ve
qual i fi cati ons may be accepted by the accr edi tati on body when staff have
extensi ve rel evant experi ence rel ati ng to the l aboratorys scope of accredi tati on.
Staff shoul d have rel evant practi cal work experi ence (at l east 2 years) before
bei ng al l owed to perform accredi ted work wi thout supervi si on or before bei ng
consi dered as experi enced for supervi si on of accredi ted work. Speci fi c nati onal
regul ati ons may overri de the gui dance gi ven i n thi s document.
3.3 The l aboratory management shoul d ensure that al l staff have recei ved adequate
trai ni ng for the competent performance of tests and operati on of equi pment.
Thi s shoul d i ncl ude trai ni ng i n basi c techni ques, eg pl ate pouri ng, counti ng of
col oni es, asepti c techni que, etc, where thi s has not previ ousl y been undertaken.
Objecti ve measures as determi ned by, eg repl i cate anal ysi s, shoul d be recorded
and used to assess the attai nment of competence duri ng trai ni ng. Staff may
onl y perform tests on sampl es i f they are ei ther recogni sed as competent to do
so, or i f they do so under adequate supervi si on. Conti nued competence shoul d
be moni tored and, where thi s i s not achi eved, the need to retrai n staff shoul d
be consi dered. Where a method or techni que i s not i n regul ar use, veri fi cati on
of staff performance before they undertake tests may be necessary. The cri ti cal
i nterval between performance of tests shoul d be establ i shed and documented.
3.4 I n some cases, i t may be more appropri ate to rel ate competence to a parti cul ar
techni que or i nstrument rather than to methods.
4 Environment
EN 45001, paragraph 5.3.2
4.1 General arrangement of the premi ses
4.1.1 There are general l y two types of premi ses i n l aboratori es; ancillary premises
(entrances, corri dors, admi ni strati on bl ocks, cl oak rooms and toi l ets, storage
rooms, archi ves, etc); and the test premises (where speci fi c mi crobi ol ogi cal
testi ng and associ ated acti vi ti es are carri ed out) for whi ch, general l y, there are
speci fi ed envi ronmental requi rements.
4.1.2 The l aboratory must be arranged so as to mi ni mi se ri sks of cross-contami nati on,
where thi s i s si gni fi cant to the type of test bei ng performed. The ways to achi eve
thi s objecti ve are, for exampl e:
(a) to construct the l aboratory to the no way back l ayout pri nci pl e;
(b) to carry out procedures i n a sequenti al manner usi ng appropri ate precauti ons
to ensure test and sampl e i ntegri ty (eg use of seal ed contai ners);
(c) to segregate acti vi ti es by ti me or space.
4.1.3 I t i s general l y consi dered as good practi ce to have separate l ocati ons, or cl earl y
desi gnated areas, for the fol l owi ng:
sampl e recei pt and storage areas;
sampl e pr epar ati on (eg a segr egated l ocati on shoul d be used for the
preparati on of powdery products l i kel y to be hi ghl y contami nated);
exami nati on of sampl es, i ncl udi ng i ncubati on;
mai ntenance of reference organi sms;
medi a and equi pment preparati on, i ncl udi ng steri l i zati on;
steri l i ty assessment;
decontami nati on.
The area for washi ng (after decontami nati on) may be shared wi th other parts
of the l aboratory provi di ng that the necessary precauti ons are taken to prevent
transfer of traces of substances whi ch coul d adversel y affect mi crobi al growth.
The need for physi cal separati on shoul d be judged on the basi s of the acti vi ti es
speci fi c to the l aboratory (eg number and type of tests carri ed out).
4.2 Envi ronment and moni tori ng
4.2.1 Laboratori es must be aware of the potenti al for contami nati on of areas both
i nsi de and outsi de the l aboratory, and shoul d demonstrate that they have taken
appropri ate measures to avoi d any such occurrence. For exampl e, the l aboratory
may need to construct physi cal barri ers to i sol ate the test premi ses.
4.2.2 The envi ronment wi thi n whi ch the mi crobi ol ogi cal anal yses are carri ed out shal l
be such that r esul ts ar e not i nval i dated. Dependi ng on the type of testi ng
acti vi ti es carri ed out, the l aboratory shal l defi ne and document the parti cul ar
arrangements i n pl ace for mi ni mi si ng the ri sks of contami nati on.
4.2.3 The l abor ator y shoul d devi se an appr opr i ate envi r onmental moni tor i ng
pr ogr amme i ncl udi ng, for exampl e, use of ai r settl ement pl ates and sur face
swabbi ng to measure trends i n l evel s of contami nati on for each type of testi ng
bei ng carri ed out. Acceptabl e background counts shoul d be assi gned and there
shoul d be a documented procedure for deal i ng wi th si tuati ons i n whi ch these
l i mi ts are exceeded.
4.2.4 Space shoul d be suffi ci ent to al l ow work areas to be kept cl ean and ti dy. The
space requi red shoul d be commensurate wi th the vol ume of anal yses handl ed
and the overal l i nternal organi sati on of the l aboratory.
4.2.5 Workrooms shoul d be appropri atel y venti l ated. Thi s may be done by natural or
forced venti l ati on, or by the use of an ai r condi ti oner. Where ai r condi ti oners
ar e used, fi l ter s shoul d be appr opr i ate, i nspected, mai ntai ned and r epl aced
accordi ng to the type of work bei ng carri ed out.
4.2.6 Ther e shoul d be a ver i fi abl e system for checki ng the acceptabi l i ty of the
bi ocontami nati on of the ai r.
4.3 Access
4.3.1 Dependi ng on the type of testi ng bei ng carri ed out, access to the mi crobi ol ogi cal
l aboratory shoul d be restri cted to authori sed personnel . Where such restri cti ons
are i n force, staff shoul d be made aware of:
(a) the i ntended use of a parti cul ar area;
(b) the restri cti ons i mposed on worki ng wi thi n such areas;
(c) the reasons for i mposi ng such restri cti ons.
4.4 Hygi ene
4.4.1 Cl othi ng appr opr i ate to the type of testi ng bei ng per for med (i ncl udi ng, i f
necessary, protecti on for hai r, beard, hands, shoes, etc) shoul d be worn i n the
mi cr obi ol ogi cal l abor ator y and r emoved befor e l eavi ng the ar ea. I n many
l aboratori es a l aboratory coat may suffi ce.
4.4.2 Dependi ng on the type of the mi crobi ol ogi cal acti vi ti es bei ng undertaken, the
desi gn and fi tti ngs of the l abor ator y must be such as to mi ni mi se potenti al
contami nati on fr om the sur r oundi ngs and fr om substances handl ed i n the
l aboratory.
4.4.3 Wal l s, fl oors, cei l i ngs and work surfaces shoul d be non-absorbent and easy to
cl ean and di si nfect. Wooden surfaces of fi xtures and fi tti ngs shal l be adequatel y
seal ed. Measures shoul d be taken to avoi d accumul ati on of dust, by the provi si on
of suffi ci ent storage space, by havi ng mi ni mal paperwork i n the l aboratory and
by prohi bi ti ng pl ants and personal possessi ons from the l aboratory work area.
The documented cl eani ng programme for l aboratory fi xtures, equi pment and
surfaces shoul d take i nto account the resul ts of envi ronmental moni tori ng and
the possi bi l i ty of cross-contami nati on.
4.4.4 Reducti on of contami nati on can be achi eved by havi ng:
smooth surfaces on wal l s, cei l i ngs, fl oors and benches (the smoothness of a
sur face i s j udged on how easi l y i t may be cl eaned). Ti l es ar e not
recommended as bench coveri ng materi al ;
concave joi nts between the fl oor, wal l s and cei l i ng;
mi ni mal openi ng of wi ndows and doors whi l e tests are bei ng carri ed out;
sun shades pl aced on the outsi de;
easy access for cl eani ng of i nternal sun shades i f i t i s i mpossi bl e to fi t them
outsi de;
fl ui d conveyi ng pi pes not passi ng above wor k sur faces unl ess pl aced i n
hermeti cal l y seal ed casi ngs;
a dust-fi l tered ai r i nl et for the venti l ati on system;
separate hand-washi ng arrangements, preferabl y non-manual l y control l ed;
cupboards up to the cei l i ng;
no rough and bare wood;
stored i tems and equi pment arranged to faci l i tate easy cl eani ng;
no furni ture, documents or other i tems other than those stri ctl y necessary
for testi ng acti vi ti es.
Thi s l i st i s not exhausti ve, and not al l exampl es wi l l appl y i n every si tuati on.
Cei l i ngs, i deal l y, shoul d have a smooth surface wi th fl ush l i ghti ng. When thi s
i s not possi bl e (as wi th suspended cei l i ngs and hangi ng l i ghts), the l aboratory
shoul d have documented evi dence that they control any resul ti ng ri sks to hygi ene
and have effecti ve means of over comi ng them, eg a sur face-cl eani ng and
i nspecti on programme.
4.4.5 I n order to faci l i tate cl eani ng, tel ephones and computers used i nsi de the test
ar ea may be fi tted wi th fl exi bl e, easy-to-cl ean tr anspar ent pr otecti on. The
computer equi pment venti l ati on system shoul d be ar r anged to pr event
contami nati on, i e the ai r fl ow shoul d not be di rected onto the workbenches.
4.4.6 I n cases wher e wor k under ster i l e condi ti ons i s l i mi ted or takes pl ace onl y
occasi onal l y, i t may be suffi ci ent to use a cl ean bench provi ded that stri ngent
asepti c techni ques are used.
5 Equipment maintenance, calibration and
performance verification
EN45001 paragraph 5.3.3
ISO Guide 25, paragraphs 7 & 9
5.1 As part of i ts qual i ty system, a l aboratory i s requi red to operate a documented
programme for the mai ntenance, cal i brati on and performance veri fi cati on of i ts
equi pment.
5.2 Mai ntenance
(Gui dance on mai ntenance of equi pment can be found i n I SO 7218.)
5.2.1 Mai ntenance of essenti al equi pment shal l be carri ed out at speci fi ed i nterval s
as determi ned by factors such as the rate of use. Detai l ed records shal l be kept.
5.2.2 Attenti on shoul d be pai d to the avoi dance of cross-contami nati on ari si ng from
equi pment, eg:
di sposabl e equi pment shoul d be cl ean and steri l e;
re-used gl assware shoul d be properl y cl eaned;
i deal l y, l aboratori es shoul d have more than one autocl ave. However, one
autocl ave i s acceptabl e provi ded that adequate precauti ons are taken to
separate decontami nati on and steri l i sati on l oads, and a documented cl eani ng
programme i s i n pl ace to address both the i nternal and external envi ronment
of the autocl ave.
5.2.3 Typi cal l y, the fol l owi ng i tems of equi pment wi l l be mai ntai ned by cl eani ng and
ser vi ci ng, i nspecti ng for damage, gener al ver i fi cati on and, wher e r el evant,
steri l i si ng:
general servi ce equi pment - fi l trati on apparatus, gl ass or pl asti c contai ners
(bottl es, test tubes), gl ass or pl asti c Petri di shes, sampl i ng i nstruments, wi res
or l oops of pl ati num, ni ckel /chromi um or di sposabl e pl asti c;
water baths, i ncubators, mi crobi ol ogi cal cabi nets, autocl aves, homogeni sers,
fri dges, freezers;
vol umetri c equi pment - pi pettes, automati c di spensers, spi ral pl aters;
measuri ng i nstruments - thermometers, ti mers, bal ances, pH meters, col ony
counters.
5.3 Cal i brati on and performance veri fi cati on
5.3.1 The l aboratory must establ i sh a programme for the cal i brati on and performance
veri fi cati on of equi pment whi ch has a di rect i nfl uence on the test resul ts. The
frequency of such cal i brati on and performance veri fi cati on wi l l be determi ned
by documented exper i ence and wi l l be based on need, type and pr evi ous
performance of the equi pment. I nterval s between cal i brati on and veri fi cati on
shal l be shorter than the ti me the equi pment has been found to take to dri ft
outsi de acceptabl e l i mi ts. Exampl es of cal i br ati on i nter val s and typi cal
performance checks for vari ous l aboratory i nstruments are gi ven i n Appendi x E
and Appendi x F.
5.3.2 Where the concept i s appl i cabl e, al l measurements havi ng a si gni fi cant effect
upon test r esul ts must be tr aceabl e to nati onal or i nter nati onal standar ds.
Evi dence of tr aceabi l i ty shal l be thr ough cer ti fi cates i ssued by a nati onal
standards l aboratory or by a l aboratory accredi ted for cal i brati on. I n the case
where traceabi l i ty i s not avai l abl e through ei ther of these routes, i t shal l be
pr ovi ded by a body r ecogni sed by the accr edi tati on or gani sati on for the
measurement concerned.
5.3.3 Temperature measurement devices
(a) Where the accuracy of temperature measurement has a di rect effect on the
r esul t of an anal ysi s, temper atur e measur i ng devi ces, eg l i qui d-i n-gl ass
thermometers, thermocoupl es, pl ati num resi stance thermometers (PRTs)
used i n i ncubators and autocl aves shal l be of the appropri ate qual i ty to
achi eve the speci fi cati on i n the test method. The gr aduati on of the
temper atur e measur i ng devi ces must be appr opr i ate for the r equi r ed
accur acy of measur ement. They shal l al so be cal i br ated to nati onal or
i nternati onal standards for temperature.
(b) Acceptabl e traceabi l i ty of measurement for thermometers may be achi eved
by i n-house cal i br ati on agai nst cal i br ated r efer ence ther mometer s,
thermocoupl es or pl ati num resi stance thermometers i n accordance wi th a
documented procedure, provi ded that the overal l uncertai nty of measure-
ment of the reference devi ce i s appropri ate to the cal i brati on.
(c) When the accuracy of the temperature measurement does not have a di rect
effect on the test resul t, eg i n the case of fri dges and freezers, l aboratori es
may meet accredi tati on requi rements by usi ng, throughout the l aboratory,
worki ng thermometers manufactured to acceptabl e nati onal /i nternati onal
speci fi cati ons. Veri fi cati on of the performance of these devi ces wi l l need to
be carri ed out.
(d) An i ndependent veri fi cati on of the i ntegral thermometers and recorders i n
medi a preparators and autocl aves shal l be carri ed out to demonstrate thei r
accuracy. Where i t i s not possi bl e to use devi ces such as thermocoupl es,
maxi mum ther mometer s that have been cal i br ated wi thi n the r equi r ed
temper atur e r ange may be used to moni tor the autocl ave chamber
temperatures. A compari son between temperatures i ndi cated external l y and
the maxi mum reached i nsi de the autocl ave may be made. Records of such
checks and detai l s of any correcti ve acti on taken shal l be recorded.
5.3.4 The stabi l i ty of temperature, uni formi ty of temperature di stri buti on and ti me
requi red to achi eve equi l i bri um condi ti ons i n i ncubators, water baths, ovens
and temperature-control l ed rooms shal l be establ i shed i ni ti al l y and documented,
i n parti cul ar wi th respect to typi cal uses (for exampl e posi ti on, space between,
and hei ght of, stacks of Petr i di shes). The constancy of the char acter i sti cs
r ecor ded dur i ng i ni ti al ver i fi cati on of the equi pment shal l be checked and
recorded after each si gni fi cant repai r or modi fi cati on. Laboratori es shal l moni tor
and retai n temperature records of equi pment used for testi ng.
5.3.5 Autoclaves
(a) Autocl aves shal l be capabl e of meeti ng speci fi ed temperature tol erances.
Pressure cookers fi tted onl y wi th a pressure gauge are not recommended
for steri l i sati on of medi a or decontami nati on of wastes.
(b) The per for mance of each autocl ave shal l be i ni ti al l y eval uated by
establ i shi ng i ts functi onal properti es, eg heat di stri buti on characteri sti cs
wi th respect to typi cal uses. Thi s process must be repeated after si gni fi cant
r epai r or modi fi cati on (eg r epl acement of ther mo-r egul ator pr obe or
pr ogr ammer , l oadi ng ar r angements, oper ati ng cycl e). The ster i l i sati on/
decontami nati on cycl e must take account of the heati ng profi l e of the l oad.
Cl ear operati ng i nstructi ons shal l be gi ven based on the heati ng profi l es
determi ned for typi cal uses.
(c) Records of autocl ave operati ons, i ncl udi ng temperature and ti me, shal l be
mai ntai ned. Thi s shoul d be done for every cycl e, acceptance/rejecti on cri teri a
set and r ecor ds mai ntai ned. Moni tor i ng shal l be achi eved by one of the
fol l owi ng:
(i ) usi ng a thermocoupl e and recorder to produce a chart or pri ntout;
(i i ) usi ng a maxi mum thermometer;
(i i i ) di rect observati on and recordi ng of maxi mum temperature achi eved.
I n addi ti on to di r ectl y moni tor i ng the temper atur e of an autocl ave, the
effecti veness of i ts operati on duri ng each cycl e may be checked by the use
of chemi cal or bi ol ogi cal i ndi cator s for ster i l i sati on/decontami nati on
purposes. Autocl ave tape shoul d be used to i ndi cate that a l oad has been
processed, but not as an i ndi cator to demonstrate compl eti on of an acceptabl e
steri l i sati on cycl e.
5.3.6 Wei ghts shal l be cal i br ated and bal ances ver i fi ed at r egul ar i nter val s by a
documented procedure (accordi ng to thei r i ntended use). Al l wei ghts shal l be
cal i brated and traceabl e to nati onal or i nternati onal standards.
5.3.7 (a) Vol umetr i c equi pment such as automati c di spenser s, di spenser /di l uter s,
mechani cal hand pi pettes and di sposabl e pi pettes may al l be used i n the
mi crobi ol ogy l aboratory. Laboratori es shoul d carry out i ni ti al veri fi cati on
of vol umetri c equi pment and then make regul ar checks to ensure that the
equi pment i s per for mi ng wi thi n the r equi r ed speci fi cati on. Ver i fi cati on
shoul d not be necessary for gl assware whi ch has been certi fi ed to a speci fi c
tol erance.
(b) Equi pment shoul d be ver i fi ed for the accur acy of the del i ver ed vol ume
agai nst the set vol ume (for several di fferent setti ngs i n the case of vari abl e
vol ume i nstruments) and the preci si on of the repeat del i veri es shoul d be
measur ed. For one-use di sposabl e vol umetr i c equi pment, l abor ator i es
shoul d obtai n suppl i es from compani es wi th rel evant I SO 9000 regi strati on.
After i ni ti al veri fi cati on of the sui tabi l i ty of the equi pment, i t i s recommended
that random checks on accuracy are carri ed out. Where compani es do not
have I SO 9000 r egi str ati on, l abor ator i es shoul d check each batch of
equi pment for sui tabi l i ty.
5.3.8 Conducti vi ty meters, oxygen meters, pH meters and other si mi l ar i nstruments
shal l be veri fi ed regul arl y or before each use. The buffers used to cal i brate the
i nstrument shal l be stored i n appropri ate condi ti ons and shal l be marked wi th
an expi ry date.
5.3.9 Where humi di ty i s i mportant to the outcome of the test, hygrometers shal l be
cal i brated and the cal i brati on shal l be traceabl e to nati onal or i nternati onal
standards.
5.3.10 Ti mers, i ncl udi ng the autocl ave ti mer, shal l be veri fi ed usi ng a cal i brated ti mer
or nati onal ti me si gnal .
6 Reagents and culture media
ISOGuide 25, paragraph 8.1
6.1 The l aboratory shoul d ensure that the qual i ty of the reagents used i s appropri ate
for the tests concerned. Preferabl y, reagents (i ncl udi ng ready-to-use medi a and
Petr i di shes) shoul d be obtai ned fr om manufactur er s who have a qual i ty
management system cer ti fi ed to I SO 9000. Labor ator i es shoul d ensur e that
certi fi cati on covers al l rel evant acti vi ti es i ncl udi ng suppl y/del i very, where thi s
has a beari ng on the performance of the goods. Laboratori es shoul d i ni ti al l y
ver i fy the sui tabi l i ty of the pr oduct by usi ng posi ti ve and negati ve contr ol
organi sms whi ch are traceabl e to recogni sed nati onal cul ture col l ecti ons.
6.2 Cul ture medi a may be prepared i n the l aboratory from the i ndi vi dual chemi cal s,
from commerci al l y avai l abl e dehydrated powders, or may be purchased ready
to use.
6.3 Reagents and commerci al dehydrated powders shal l be consumed wi thi n the
shel f-l i fe of the product. The date of recei pt, expi ry date and openi ng date shoul d
be recorded. The stock shoul d be rotated so that the ol der medi a and reagents
are used fi rst. Storage shoul d be under appropri ate condi ti ons, eg cool , dry and
dark. Al l contai ners, especi al l y those for dehydrated medi a, shoul d seal ti ghtl y.
Dehydrated medi a that are caked or cracked or show a col our change shoul d
not be used.
6.4 Laboratory prepared medi a, sol uti ons and buffers shoul d, general l y, be made
from bacteri ol ogi cal grade chemi cal s.
6.5 Where l aboratori es are maki ng use of pre-prepared medi a and reagents, they
shoul d obtai n a copy of the I SO 9000 regi strati on certi fi cate from the suppl i ers
of the goods. I t i s recommended that further checks shoul d be made on products
on a r andom basi s to ensur e conti nued compl i ance wi th the r equi r ed
speci fi cati on. These checks may be encompassed by a l abor ator ys i n-house
regul ar QC programme. The manufacturer shoul d i ni ti al l y suppl y a qual i ty
speci fi cati on whi ch wi l l i ncl ude at l east the fol l owi ng:
(a) shel f l i fe of the product;
(b) storage condi ti ons;
(c) sampl i ng regi me/rate;
(d) steri l i ty check, i ncl udi ng acceptabi l i ty cri teri a;
(e) effi cacy checks i ncl udi ng the organi sm used, thei r cul ture col l ecti on reference
and acceptabi l i ty cri teri a;
(f) date of i ssue of speci fi cati on.
Each batch recei ved shoul d i ncl ude an assurance that i t i s suppl i ed i n accordance
wi th the qual i ty speci fi cati on. I n the event of any changes, the manufacturer
shoul d suppl y a revi sed speci fi cati on.
6.6 Di sti l l ed water, de-i oni sed water or reverse osmosi s produced water, free from
bacteri ci dal and i nhi bi tory substances, shoul d be used i n the preparati on of
medi a, sol uti ons and buffers.
6.7 Medi a, sol uti ons and reagents shoul d be prepared, used and stored i n accordance
wi th a documented procedure fol l owi ng the i nstructi ons of the manufacturer/
author . Gui dance on the pr epar ati on and ster i l i sati on of medi a, and
recommended storage ti mes can be found i n I SO 7218.
6.8 Al l l abor ator y pr epar ed batches of medi a shoul d be checked to ensur e they
support the growth of speci fi c mi crobi al cul tures. I n addi ti on, sel ecti ve medi a
shoul d be checked to ensure they suppress the growth of non-target organi sms.
I n preference to usi ng the commonl y used streak method, i t i s better to use a
quanti tati ve procedure, where a known (often l ow) number of rel evant organi sms
are i nocul ated onto the medi um under test and the recovery eval uated. Thi s
can be used to establ i sh a recovery l evel bel ow whi ch a batch wi l l not be accepted.
6.9 Labor ator i es shal l ensur e that al l r eagents (i ncl udi ng stock sol uti ons) ar e
adequatel y l abel l ed to i ndi cate i denti ty, concentrati on, storage condi ti ons, expi ry
date and/or r ecommended stor age per i ods. The per son r esponsi bl e for the
preparati on of the reagent shoul d be i denti fi abl e ei ther from the l abel or from
the records.
7 Test methods and procedures
EN 45001, paragraph 5.4
7.1 Laboratori es may use sectoral , offi ci al , nati onal , and i nternati onal standard
methods and i n-house methods. The l aboratory shoul d not feel constrai ned to
use a standard method i f i t has an i n-house method whi ch has equi val ent or
super i or per for mance, mor e moder n technol ogy and a degr ee of val i dati on
adequate for the purpose. The l aboratory shoul d sati sfy i tsel f that each parti cul ar
method i s adequate for i ts i ntended purpose
7.2 The tr ueness, r epeatabi l i ty/r epr oduci bi l i ty, speci fi ci ty, sensi ti vi ty, l i mi t of
determi nati on, matri x effects and ease of use must be taken i nto account before
sel ecti ng a parti cul ar test method. Laboratori es shoul d sel ect methods whi ch
are sui tabl e for thei r purposes (see Secti on 9).
7.3 Methods used by a l aboratory shal l be ful l y documented. A recommendati on for
these procedures i s gi ven i n I SO 78/2, Layouts for Standards: Part 2.
8 Validation of methods and verification of
performance
8.1 Each l abor ator y wi l l have par ti cul ar r equi r ements for the per for mance
characteri sti cs of a parti cul ar method i n order to demonstrate sui tabi l i ty for
the i ntended purpose. However, the essenti al feature of any method i s that i t
shoul d gi ve the correct answer wi th respect to speci fi ed l i mi ts of detecti on,
sel ecti vi ty, repeatabi l i ty and reproduci bi l i ty.
8.2 For offi ci al methods, or methods fr om r ecogni sed nati onal or i nter nati onal
standard organi sati ons, a ful l val i dati on may not be necessary but, before usi ng
such a method for the fi r st ti me, the l abor ator y shoul d i ntr oduce i t by a
documented trai ni ng programme. Basi c parameters l i ke vari ati on, sel ecti vi ty,
sensi ti vi ty and speci fi ci ty can general l y be found i n sci enti fi c publ i cati ons, books
and manual s for mi crobi ol ogi cal medi a.
8.3 Commer ci al i sed test systems (ki ts) may not r equi r e fur ther val i dati on i f
val i dati on data from al ternati ve sources, eg based on col l aborati ve testi ng, i s
avai l abl e. Laboratori es shoul d seek from manufacturers val i dati on data and
evi dence of oper ati on to a r ecogni sed qual i ty assur ance system. Wher e ful l
val i dati on data are not avai l abl e, the l aboratory i s responsi bl e for compl eti ng
the val i dati on of the method before usi ng i t routi nel y.
8.4 For al l other methods, val i dati on must be performed to assure the rel i abi l i ty of
the obtai ned resul ts and, i f possi bl e, to establ i sh the resul ts di spersi on.
8.5 Qual i tati ve mi crobi ol ogi cal test methods (i n whi ch the response i s expressed i n
terms of presence/absence) shoul d be val i dated by esti mati ng , i f appropri ate,
the speci fi ci ty, rel ati ve trueness, posi ti ve devi ati on, negati ve devi ati on, l i mi t of
detecti on, matri x effect, repeatabi l i ty and reproduci bi l i ty (see Appendi x A for
defi ni ti ons).
8.6 For quanti tati ve mi crobi ol ogi cal test methods, the speci fi ci ty, sensi ti vi ty, rel ati ve
trueness, posi ti ve devi ati on, negati ve devi ati on, repeatabi l i ty, reproduci bi l i ty
and the l i mi t of determi nati on wi thi n a defi ned vari abi l i ty shoul d be consi dered
and, i f necessary, quanti tati vel y determi ned i n assays. The di fferences due to
the matri ces must be taken i nto account when testi ng di fferent types of sampl es.
The resul ts shoul d be eval uated wi th appropri ate stati sti cal methods.
8.7 The val i dati on of mi crobi ol ogi cal test methods shoul d be performed under the
same condi ti ons as those of a r eal assay. Thi s can be achi eved by usi ng a
combi nati on of natural l y contami nated products and spi ked products.
8.8 Al l val i dati on data must be recorded and stored for at l east as l ong as the method
i s i n force and as l ong as necessary to ensure adequate traceabi l i ty of raw data
and resul ts.
8.9 Parti ci pati on i n, or organi sati on of, col l aborati ve tri al s, profi ci ency testi ng, or
i nter-l aboratory compari sons, whether formal or i nformal , i s al so a means of
checki ng the val i di ty of methods but i t i s recogni sed that thi s i s not al ways
feasi bl e. The anal ysi s of sampl es usi ng both the pr oposed new method and
exi sti ng methods for the same determi nati on woul d assi st i n establ i shi ng the
effi cacy of a method.
8.10 I f a modi fi ed versi on of a method i s requi red to meet the same speci fi cati on as
the ori gi nal method, then compari sons shoul d be carri ed out usi ng repl i cates to
ensure that thi s i s the case. A stati sti cal l y acceptabl e number of sampl es shoul d
be anal ysed by each procedure to ascertai n whether any di fference i n the resul ts
i s stati sti cal l y si gni fi cant.
8.11 Even when val i dati on i s compl ete, the user wi l l sti l l need to veri fy that the
documented performance can be met, eg by the use of spi ked sampl es.
9 Quality assurance of results/quality control
EN45001 paragraph 5.4.2 (e) and (f)
The I SO defi ni ti on for qual i ty assurance i s gi ven i n Appendi x A.
9.1 Qual i ty assurance i s the programme of acti vi ti es carri ed out by a l aboratory
i ntendi ng to i mprove l aboratory performance general l y. The acti vi ti es i ncl ude
encouragement of the constant use of i nternal qual i ty control , support of external
assessment schemes, and al l measur es taken to i ncr ease both wi thi n and
between l aboratory reproduci bi l i ty by means of trai ni ng courses, conferences,
and col l aborati ve studi es of l aboratory methods.
9.2 I nternal qual i ty control
9.2.1 I nternal qual i ty control consi sts of the procedures undertaken by a l aboratory
for the conti nual eval uati on of the work of the l aboratory. The mai n objecti ve
i s to ensure that day-to-day consi stency of measurement i s i n agreement wi th
some agreed val ue, such as by compari son wi th the agreed characteri sti cs of
mol ecul es, cel l s, organi sms or wi th the assi gned val ues of control materi al s where
these exi st. When consi stency i s not achi eved, control must be exerci sed over
the rel ease of resul ts.
9.2.2 Laboratori es shoul d operate i nternal qual i ty control schemes usi ng, whenever
appropri ate, stati sti cal techni ques such as:
desi gn of experi mental /factori al anal yses;
vari ati on/regressi on anal yses;
safety eval uati on/ri sk anal ysi s;
tests of si gni fi cance;
qual i ty control charts;
stati sti cal sampl i ng i nspecti on.
9.3 Reference Cul tures
9.3.1 To demonstrate traceabi l i ty, l aboratori es shal l use reference cul tures of mi cro-
organi sms obtai ned from a recogni sed nati onal col l ecti on or an organi sati on
recogni sed by the accredi tati on body.
9.3.2 Reference cul tures may be sub-cul tured once to provi de reference stocks. Puri ty
and bi ochemi cal checks shoul d be made as appropri ate. The reference stocks
shal l be preserved by a techni que (eg, freeze-dryi ng, l i qui d ni trogen storage,
deep-fr eezi ng) whi ch mai ntai ns the desi r ed char acter i sti cs of the str ai ns.
Reference stocks shal l be used to prepare worki ng stocks for routi ne work (see
Appendi x D on preparati on of bacteri al worki ng stocks). I f reference stocks have
been thawed, they must not be re-frozen and re-used.
9.3.3 Bacteri al worki ng stocks shoul d not normal l y be sub-cul tured. However worki ng
stocks may be sub-cul tured up to a defi ned number of sub-cul tures when:
(a) i t i s requi red by standard methods; or
(b) l aboratori es can provi de documentary evi dence demonstrati ng that there
has been no l oss of vi abi l i ty (where i mportant), no changes i n bi ochemi cal
acti vi ty and/or no change i n morphol ogy.
Worki ng stocks shal l not be sub-cul tured to repl ace reference stocks.
9.4 Reference materi al s and certi fi ed reference materi al s
9.4.1 Refer ence mater i al s and cer ti fi ed r efer ence mater i al s (see defi ni ti on i n
Appendi x A) provi de essenti al traceabi l i ty i n measurements and are used, for
exampl e, to demonstr ate the accur acy of r esul ts, cal i br ate equi pment and
methods, moni tor l aboratory performance and val i date methods, and enabl e
compari son of methods by use as transfer standards. Thei r use i s encouraged
wherever possi bl e.
9.4.2 Reference materi al s and substances and certi fi ed reference materi al s shal l be
stor ed and handl ed under condi ti ons that do not al ter thei r i ntegr i ty, i n
accordance wi th a documented procedure and the rel evant test method.
9.5 External qual i ty assessment (profi ci ency testi ng)
9.5.1 External l y organi sed profi ci ency testi ng schemes provi de an i ndependent means
by whi ch a l aboratory may objecti vel y assess and demonstrate the rel i abi l i ty of
resul ts produced by i ts anal yti cal methods. Parti ci pati on provi des a means for
a l aboratory to measure i ts own performance agai nst that of other l aboratori es.
I t i s i mportant to moni tor profi ci ency testi ng resul ts as a means of checki ng
qual i ty assurance and to take appropri ate acti on as necessary.
9.5.2 Accr edi tati on bodi es r ecogni se the benefi t of these schemes and str ongl y
encourage l aboratori es to parti ci pate i n profi ci ency testi ng as an i mportant part
of thei r qual i ty assurance protocol s. Laboratori es shoul d regul arl y parti ci pate
i n schemes whi ch are rel evant to thei r scope of accredi tati on; i n speci fi c i nstances
parti ci pati on may be a requi rement. I nformati on on rel evant schemes may be
provi ded by the accredi tati on organi sati on.
10 Laboratory audit and review
EN45001, paragraph 5.4.2
ISOGuide 25, paragraphs 5.3-5.5
EAL Information Sheet, EAL-G3
10.1 Audi t and revi ew are i mportant aspects i n the operati on of a qual i ty system.
Quality audit i s the peri odi c check that a l aboratory organi ses i ts own qual i ty
system to ensure that i t i s effecti ve, i mpl emented and adhered to. Quality
review i s the peri odi c exami nati on of the qual i ty system to ensure that i t meets
the needs of the l aboratory and the requi rements of the qual i ty standards.
10.2 Audi t and r evi ew ar e descr i bed i n detai l i n the Eur opean cooper ati on for
Accredi tati on of Laboratori es (EAL) publ i cati on EAL-G3, and l aboratori es must
al so ensure that speci fi c detai l s gi ven i n thi s document are i ncl uded i n the audi t.
11 Sample handling and identification
EN 45001, paragraph 5.4.5
ISOGuide 25, paragraphs 10 & 11
11.1 Sampl i ng acti vi ti es outsi de the l aboratory are not di rectl y covered by EN 45001
or I SO Gui de 25. However , mi cr obi al fl or a may be sensi ti ve to factor s l i ke
temperature or durati on of storage and transport, so i t i s i mportant to check
and record the condi ti on of the sampl e on recei pt by the l aboratory.
11.2 The l aboratory shoul d have a procedure that covers the del i very of sampl es. I f
there i s i nsuffi ci ent sampl e or the sampl e i s i n poor condi ti on due to physi cal
deteri orati on, i ncorrect temperature, torn packagi ng or defi ci ent l abel l i ng, the
l aboratory shoul d ei ther refuse the sampl e or (i f i t i s possi bl e to carry out the
work) shoul d i ndi cate the condi ti on on the test report.
11.3 The fol l owi ng i nformati on shoul d be noted:
(a) a uni que unambi guous i denti fi cati on that can be used to trace the sampl e
from recei pt to the end of the anal yti cal process;
(b) date and, where rel evant, the ti me of recei pt;
(c) i denti ty of person/organi sati on provi di ng sampl e for test;
(d) sampl e i denti fi cati on number from the sampl er (i f any);
(e) nature and characteri sti cs of the sampl e;
(f) l i st of tests requi red,
and, as far as i s necessary,
(g) temperature and condi ti on of the sampl e on recei pt;
(h) characteri sti cs of the sampl i ng operati on (sampl i ng date, sampl i ng condi ti ons
etc).
11.4 Sampl es awai ti ng test shal l be stored under sui tabl e condi ti ons to mi ni mi se
any modi fi cati ons to any mi crobi al popul ati on present.
11.5 The packagi ng and l abel s from sampl es may be hi ghl y contami nated and shoul d
be handl ed and stored wi th care so as to avoi d any spread of contami nati on.
11.6 The preparati on of the l aboratory sampl e and the test porti on shoul d fol l ow the
nati onal or i nternati onal standards speci fi c to the tested products (i f avai l abl e)
and the general gui dance gi ven i n I SO 6887 and I SO 7218.
11.7 Sampl e preparati on may si mpl y i nvol ve sti rri ng a sampl e and measuri ng an
al i quot (eg l i qui ds) or may requi re a mul ti -stage reconsti tuti on and sub-cul turi ng
(eg dri ed products). I n ei ther case the l aboratory shoul d be abl e to demonstrate
that:
(a) the test porti on i s as representati ve of the product as possi bl e (when rel evant)
and sui tabl e for anal ysi s;
(b) contami nati on of the test porti on and the envi ronment has been avoi ded
(see Secti on 4).
11.8 A pr ocedur e for the r etenti on and di sposal of sampl es shal l be wr i tten.
Laboratory sampl e porti ons that are known to be hi ghl y contami nated shal l be
decontami nated pri or to bei ng di scarded. They shoul d be stored unti l the test
resul ts are obtai ned, or l onger i f necessary.
12 Disposal of contaminated waste
12.1 The correct di sposal of contami nated materi al s may not di rectl y affect the qual i ty
of sampl e anal ysi s, however i t i s a matter of good l aboratory management and
shoul d conform to nati onal /i nternati onal envi ronmental or heal th and safety
regul ati ons (see al so I SO 7218).
13 Uncertainty of measurement
13.1 The i nter nati onal defi ni ti on for uncer tai nty of measur ement i s gi ven i n
I SO I nternati onal vocabul ary of basi c and general terms i n metrol ogy : 1993
(see Appendi c C).
13.2 I t i s recogni sed that the current state of knowl edge regardi ng uncertai nty of
measurement across the ful l range of mi crobi ol ogi cal di sci pl i nes i s vari abl e. For
thi s reason, l aboratori es may not currentl y have access to appropri ate gui dance
on esti mati ng uncertai nty of measurement i n thei r parti cul ar di sci pl i ne. Thi s
si tuati on i s currentl y bei ng addressed wi thi n the l aboratory communi ty and i t
i s expected that more cl earl y defi ned gui dance i n the fi el d of mi crobi ol ogy wi l l
be avai l abl e i n futur e. However , r epeatabi l i ty and r epr o-duci bi l i ty data ar e
components of uncertai nty of measurement and shoul d be determi ned as a fi rst
step towards produci ng esti mates of thi s parameter.
14 Use of computers
EN45001, paragraphs 5.3.3, 5.4
14.1 Where wel l establ i shed software i s used for the purpose of communi cati on or
anal yti cal work, no parti cul ar val i dati on i s necessary.
14.2 Where i n-house software i s used, compl ete documentati on for val i dati on purposes
must be provi ded. Furthermore, i t must be shown that l oss or corrupti on of data
does not occur.
14.3 Where software i s updated, a record of the revi si ons must be retai ned.
material
Appendix A
Gl ossary of Terms
Certified reference Reference materi al , accompani ed by a certi fi cate, one or
mor e of whose pr oper ty val ues ar e cer ti fi ed by a
procedure, whi ch establ i shes traceabi l i ty to an accurate
real i sati on of the uni t i n whi ch the property val ues are
expr essed, and for whi ch each cer ti fi ed val ue i s
accompani ed by an uncer tai nty at a stated l evel of
confi dence.
[I SO Gui de 30:1992, Terms and defi ni ti ons used i n
connection with reference materials]
Limit of The l owest number of organi sms wi thi n a defi ned vari -
abi l i ty that may be determi ned under the experi mental
condi ti ons of the method under eval uati on.
Limit of detection The l owest number of mi cr o-or gani sms that can be
detected, but i n number s that cannot be esti mated
accuratel y.
Linearity The abi l i ty of the method, when used wi th a gi ven
matr i x, to gi ve r esul ts that ar e i n pr opor ti on to the
number s of mi cr o-or gani sms that ar e pr esent i n the
sampl e.
Negative deviation Occurs when the al ternati ve method gi ves a negati ve
resul t wi thout confi rmati on when the reference method
gi ves a posi ti ve resul t. Thi s devi ati on becomes a fal se
negati ve resul t when the true resul t can be proved as
bei ng posi ti ve.
Positive deviation Occur s when the al ter nati ve method gi ves a posi ti ve
resul t wi thout confi rmati on when the reference method
gi ves a negati ve resul t. Thi s devi ati on becomes a fal se
posi ti ve resul t when the true resul t can be proved as
bei ng negati ve.
Quality assurance Al l those pl anned and systemati c acti ons necessary to
provi de adequate confi dence that a product or servi ce wi l l
sati sfy gi ven requi rements for qual i ty. [I SO 8402:1986
Qual i ty vocabul ary]
Quality control The operati onal techni ques and acti vi ti es that are used
to ful fi l requi rements for qual i ty.
[I SO 8402:1986, Quality vocabulary]
Reference culture Cul tures of mi cro-organi sms obtai ned from a recogni sed
nati onal col l ecti on.
determination
Reference material Mater i al or substance one or mor e of whose pr oper ty
val ues are suffi ci entl y homogeneous and wel l establ i shed
to be used for the cal i br ati on of an appar atus, the
assessment of a measurement method, or for assi gni ng
val ues to materi al s.
[I SO Gui de 30:1992, Terms and definitions used in
connection with reference materials]
Reference method Thor oughl y i nvesti gated method, cl ear l y and exactl y
descri bi ng the necessary condi ti ons and procedures, for
the measurement of one or more property val ues that
has been shown to have accur acy and pr eci si on
commensur ate wi th i ts i ntended use and that can
therefore be used to assess the accuracy of other methods
for the same measurement, parti cul arl y i n permi tti ng
the characteri sati on of a reference materi al .
Reference stocks Reference cul tures hel d by a l aboratory.
Relative trueness The degree of correspondence of the resul ts of the method
under eval uati on to those obtai ned usi ng a recogni sed
reference method, such as those provi ded by Nati onal
Standar ds Or gani sati on, the I nter nati onal Standar ds
Organi sati on (I SO), or by a sui tabl e trade organi sati on
I nternati onal Dai ry Federati on (I DF).
Repeatability Cl oseness of the agr eement between the r esul ts of
successi ve measurements of the same measurand under
the same condi ti ons of measurement.
[I SO I nternational vocabulary of basic and general terms
in metrology: 1993]
Reproducibility Cl oseness of the agr eement between the r esul ts of
measurements of the same measurand carri ed out under
changed condi ti ons of measurement.
[I SO I nternational vocabulary of basic and general terms
in metrology: 1993]
Sensitivity The abi l i ty of the method to detect sl i ght vari ati ons i n
the number of mi cro-organi sms wi thi n a gi ven matri x.
Specificity The degree to whi ch a method i s affected by the other
components i n a mul ti -component sampl e. A speci fi c
method i s one that i s not affected by any of the other
components.
Working stocks Sub-cul tures of mi cro-organi sms deri ved from reference
stocks to be used on a day-to-day basi s.
Verification Confi rmati on by exami nati on and provi si on of evi dence
that speci fi ed requi rements have been met.
[I SO/I EC Gui de 25: 1990, General requirements for the
competence of calibration and testing laboratories, 3rd
Edi ti on]
Appendix B
Bi bl i ography
1 Dr aft I SO/DI S 7218, Mi crobi ol ogy - General rul es for mi crobi ol ogi cal
examinations [Revi si on of fi rst edi ti on I SO 7218 : 1985]
2 Worl d Heal th Organi sati on (1983), Laboratory Biosafety Manual, Geneva: WHO.
3 I SO 3534-1 : 1993, Statistics - Vocabulary and symbols - Part 1: Probability
and general statistical terms.
4 Manual of Methods for General Bacteriology, Phi l i pp Gerhard et al ., Ameri can
Soci ety for Mi crobi ol ogy, Washi ngton 1981.
Appendix C
References
1 EN 45001 : 1989, General criteria for the operation of testing laboratories.
2 I SO/I EC Gui de 25, General requirements for the competence of calibration and
testing laboratories. 3rd Edi ti on: 1990.
3 I SO 9000-1 : 1994, Quality management and quality assurance standards.
Part 1 - Guide to selection and use.
4 Dr aft I SO/DI S 7218, Mi crobi ol ogy - General rul es for mi crobi ol ogi cal
examinations [Revi si on of fi rst edi ti on I SO 7218 : 1985]
5 I SO 78/2 : 1982. Layouts for standards.
6 EAL-G3 : Jan 1996, I nternal Quality Audits and Reviews, European cooperati on
for Accredi tati on of Laboratori es.
7 I SO 6887 : 1983, Preparation of dilutions.
8 I SO Gui de 30 : 1992, Terms and definitions used in connection with reference
materials.
9. I SO 8402 :1986. Quality vocabulary.
10. I SO I nternational vocabulary of basic and general terms in metrology: 1993.
Appendix D
Use of reference cul tures (bacteri a)
Appendix E
Gui dance on cal i brati on and cal i brati on checks
Thi s i nformati on i s provi ded for gui dance purposes and the frequency wi l l be
based on the need, type and previ ous performance of the equi pment.
Type of equipment Requirement Suggested frequency
Reference Ful l traceabl e re-cal i brati on Every 5 years
thermometers & Si ngl e poi nt Annual l y
Reference (eg i ce-poi nt check)
thermocouples
Working Check agai nst r efer ence Annual l y
thermometers & thermometer at i ce-poi nt
Working and/or worki ng temperature
thermocouples r ange
Balances Ful l tr aceabl e cal i br ati on Annual l y
Calibration Ful l traceabl e cal i brati on Bi -annual l y or
weights annual l y, dependi ng
on cl ass
Check Check agai nst cal i br ated Annual l y
weight(s) wei ght or check on bal ance
i mmedi atel y fol l owi ng
traceabl e cal i brati on
Timers Check agai nst nati onal ti me Annual l y
si gnal
Volumetric Gr avi metr i c cal i br ati on to Annual l y
glassware requi red tol erance
Microscopes Traceabl e cal i brati on of I ni ti al l y
stage mi crometer
Hygrometers Tr aceabl e cal i br ati on Annual l y
Appendix F
Gui dance on commi ssi oni ng and veri fi cati on of performance
Temperature (a)Establ i sh stabi l i ty and (a)I ni ti al l y, and after
controlled equipment uni for mi ty of temper atur e r epai r /modi fi cati on
(incubators, baths,
fridges, freezers) (b)Moni tor temperature (b)Dai l y/each use
Sterilising ovens (a)Establ i sh stabi l i ty and (a)I ni ti al l y, and after
uni for mi ty of temper atur e r epai r /modi fi cati on
(b)Moni tor temperature (b)Each use
Autoclaves (a)Establ i sh characteri sti cs for (a)I ni ti al l y, and after
typi cal l oads/cycl es r epai r /modi fi cati on
(b)Moni tor temperature/ti me (b)Each use
Safety cabinets (a)Establ i sh performance (a)I ni ti al l y, and after
r epai r /modi fi cati on
(b)Par ti cl e count moni tor i ng (b)Weekl y
(c) Ai r fl ow moni tor i ng (c) Monthl y
Laminar air flow (a)Establ i sh performance (a)I ni ti al l y, and after
cabinets r epai r /modi fi cati on
(b)Check wi th ster i l i ty pl ates (b)Weekl y
Microscopes Check al i gnment Dai l y/each use
pH meters Cal i brati on check usi ng at Dai l y/each use
l east two buffers
Balances Check zero, and readi ng Dai l y/each use
agai nst check wei ght
Stills, de-ionisers (a)Check conducti vi ty (b)Dai l y
and reverse osmosis
units (b)Check for mi cr obi al (b)Monthl y
contami nati on
Gravimetric diluters (a)Check wei ght and (a)Dai l y
vol ume (wei ght) di spensed
(b)Check di l uti on r ati o (b)Monthl y
Media dispensers Check vol ume di spensed Dai l y/each use/
each adjustment
... continued
Pipettors/pipettes Check accuracy and Regul arl y (to be
preci si on of vol ume defi ned by taki ng
di spensed account of the
frequency and
natur e of use)
Spiral platers (a) Establ i sh performance (a)I ni ti al l y and
agai nst conventi onal method annual l y
(b) Check styl us condi ti on and (b)Dai l y/each use
the start and end poi nts
(c) Check vol ume di spensed (c) Monthl y
Colony counters Check agai nst number Annual l y
counted manual l y
Anaerobic Check wi th anaerobi c Each use
jars/incubators i ndi cator
Laboratory Moni tor for ai r bor ne and Weekl y
environment surface mi crobi al
contami nati on usi ng, eg
ai r sampl er s, settl e pl ates,
contact pl ates or swabs
Appendi x F (conti nued)
Appendix G
Gui dance on mai ntenance of equi pment
Incubators, fridges Cl ean and di si nfect i nter nal Monthl y
freezers, ovens sur faces
Water baths Empty, cl ean, di si nfect Monthl y, or every
and refi l l 6 months i f bi oci de
used
Centrifuges (a)Ser vi ce (a)Annual l y
(b)Cl ean and di si nfect (b)Each use
Autoclaves (a)Make vi sual checks of (a)Regul arl y, as
gasket, cl ean/drai n chamber recommended by
manufactur er
(b)Ful l ser vi ce (b)Annual l y
(c) Safety check of pr essur e (c) Annual l y
vessel
Safety cabinets Ful l servi ce and mechani cal check Every si x months
Laminar flow Servi ce and mechani cal check As recommended
cabinets by manufacturer
Microscopes (a)Cl ean, and ful l mai ntenance (a)Every 6 months
ser vi ce
(b)Check eye-pi ece grati cul e (b)Every 6 months
pH meters Cl ean el ectrode Each use
Balances, (a)Cl ean (a)Each use
gravimetric diluters
(b)Ser vi ce (b)Annual l y
Stills Cl ean and de-scal e As requi red
(eg every 3 months)
De-ionisers, Repl ace cartri dge/membrane As recommended
reverse osmosis units by manufacturer
Anaerobic jars Cl ean/di si nfect After each use
Media dispensers, Decontami nate, cl ean and Each use
volumetric equipment, steri l i se as appropri ate
pipettes, and general
service equipment
... continued
Appendi x G (conti nued)
Spiral platers (a) Ser vi ce (a)Annual l y
(b) Decontami nate, cl ean and (b)Each use
steri l i se
Laboratory (a) Cl ean and di si nfect (a)Dai l y, and duri ng
wor ki ng sur faces use
(b) Cl ean and di si nfect (b)Weekl y
fl oor s, si nks and basi ns
(c) Cl ean and di si nfect (c) Every 3 months
other surfaces

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LAB 38 Accreditation for microbiological laboratories

Edition 1 September 2001 Page A (+ pp 1 - 28)

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