65e804280dd4340bc20b10ee - Glucose SOP 6-2-22

Title: Diabetes Diagnostic Laboratory - Glucose Testing using the
Roche Cobas c311

Document Owner: Last Approved Date:
Rhonda Rojas 06/02/2021
Content Expert:
Curt Rohlfing
Printed copies are for reference only. Please refer to the electronic copy for the latest version.
I. Purpose Statement
a. This SOP is intended for the in vitro test for the quantitative determination of
glucose in human plasma. It is performed on the Roche Cobas c311 system in the
Diabetes Diagnostic Laboratory.
II. Definitions
a. Not Applicable
III. Content
a. Principle and Clinical Significance
Glucose is the major carbohydrate present in the peripheral blood. Glucose

derived from dietary sources is either oxidized to provide energy or converted to
glycogen or fatty acids for storage in the liver and tissues. The most frequent
cause of hyperglycemia is diabetes mellitus. Some other factors that contribute
to elevated blood glucose are pancreatitis, pituitary or thyroid dysfunction, renal
failure, and liver disease. Hypoglycemia is less frequently observed, but is found
in conditions such as insulinoma, hypopituitarism, neoplasms, or insulin-induced
hypoglycemia1,2,3,4.
The cobas c311 performs a UV test to detect glucose in blood serum and plasma.
The enzyme hexokinase (HK) catalyzes the reaction between glucose and
adenosine triphosphate (ATP) to form glucose-6-phosphate (G-6-P) and
adenosine diphosphate (ADP). In the presence of nicotinamide adenine
dinucleotide (NAD), G-6-P is oxidized by the enzyme glucose-6-phosphate
dehydrogenase (G-6-PD) to 6-phosphogluconate and reduced nicotinamide
adenine dinucleotide (NADH). The increase in NADH concentration is directly
proportional to the glucose concentration and can be measured
spectrophotometrically at 340 nm.
Version #: 4 Page 1 of 20
Title: Diabetes Diagnostic Laboratory - Glucose Testing using
the Roche Cobas c311
The enzymatic reference method is performed with hexokinase.4 Hexokinase
catalyzes the phosphorylation of glucose to glucose-6-phosphate by ATP.
HK
Glucose + ATP G-6-P + ADP
G-6-PDH
G-6-P + NADP+ gluconate-6-P + NADPH + H+
b. Specimen Collection and Handling

i. Patient Preparation
1. Patients should be fasting or undergoing an Oral Glucose Tolerance
Test. Patient’s status should be recorded when specimen is drawn.
ii. Specimen Type
1. Collect blood by venipuncture from individuals using an evacuated
tube system
a. Plasma: Fluoride plasma for NHANES study testing (grey top
tubes)
2. The minimum volume required for analysis directly from collection
tube is 200 µL.
3. Specimens are delivered to the Diabetes Diagnostic Laboratory,
Room M764 by FedEx. Specimens are received frozen on dry ice
and are stored at -70oC. Each specimen must arrive in the
laboratory labeled with two unique accession number generated
by NHANES, and checked against the specimen manifest.
iii. Specimen Stability

1. Stability in fluoride plasma:5 3 days at 15-25 °C
At least 3 months @ -20°C
At least 2 years @-70°C
2. Unacceptable specimen criteria

a. Clotted specimens.
b. Unlabeled samples
c. Specimens not collected in sodium fluoride.
d. Only plasma specimens are acceptable.
e. Refer to limitations section for additional details
f. The below must be followed for unaccepted specimen
collections:
i. Record all unacceptable samples on sample
manifest AND
ii. Contact NHANES ASAP regarding unacceptable
specimen.
iv. Handling Conditions

1. For specimen collection and preparation, only use suitable tubes or
collection containers. Only the specimens listed below were tested
and found acceptable: Plasma: fluoride plasma for NHANES study
testing. (Grey-top tubes)
2. To minimize glycolysis, one should place the sample tube
immediately in an ice-water slurry, and the plasma should be
separated from the cells within 30 min. Blood for FPG analysis
should be drawn in the morning after the individual has fasted
overnight (at least 8 hours).
3. If testing is not performed immediately, plasma samples are to be
maintained under frozen conditions (-70°C).
4. For NHANES samples: Laboratory services are requested through
the Westat system operations via an email notification containing
a unique manifest list of the samples and sample analysis type (e.g.
Glucose), which confirms that specimens have been shipped to
DDL. Transport under frozen conditions.
5. Each Manifest Form should include and verified against each
sample received:
a. Patient Sample ID #
b. Test Name
c. Date Collected
d. Shipment ID #
e. Shipment Date
f. Lab Name
g. Lab ID
h. Survey Year
6. Once specimens are received and verified the corresponding file is
imported electronically into the SQL server database via secure
transfer. Refer to DDL’s NHANES study sample handling and
reporting procedure for additional details.
7. After being verified specimens are to be immediately returned to -
70C storage. When ready to be analyzed, specimens are to be
thawed at room temperature (~ 35 min). Following testing,
specimens are to be returned to -70C for long-term storage.
8. Refer to the Diabetes Diagnostic Laboratory NHANES study sample
handling and reporting for additional details.
https://muhealth.policytech.com/dotNet/documents/?docid=2952
0
c. Equipment and Materials

i. Equipment
1. Roche Cobas C311 analyzer consists of the following main
components: Refer to the Operator’s Manual attached in this
procedure for START UP (Chapter 3), OPERATION (Chapter 2) and
SHUT DOWN (Chapter 8) Procedures.
a. Analyzer Unit
i. Sample disk: 110 position disk for loading of up to
108 samples
ii. Sample detectors and barcode readers: on inner and
outer rings
iii. Sample Pipetting System: clot detecting by pressure
measurements, liquid level detection by
capacitance, and abnormal descent detection.
1. Metal shield pipe
2. Rinse station
3. Drying cylinder
4. Basic (sample cleaner 1) and acid wash
(sample cleaner 2) solutions for special
probe washes and maintenance functions.
iv. Reagent Compartment
v. Reagent Pipetting System
vi. Reaction Disk Area
vii. ISE System (not in use)
viii. Water supply tank
ix. Vacuum System
x. Sound Volume Knob
xi. Cell Wash Solutions
xii. Waste Container
xiii. Inlet Water Line
xiv. Dilute Waste Line
b. Control Unit
i. Computer
1. Hard Disk Drive
2. 3-1/2 in floppy disk drive and a DVD-RW
rewrite device
ii. Keyboard
iii. Mouse
iv. Color Monitor: touch screen
v. Printer
ii. Materials
1. Reagents and other items
a. R1: MES buffer: 5.0 mmol/L, pH 6.0; Mg2+: 24 mmol/L;
ATP:  4.5 mmol/L; NADP:  7.0 mmol/L; preservative
b. R2: HEPES buffer: 200 mmol/L, pH 8.0; Mg2+: 4 mmol/L; HK
(yeast):  300 µkat/L; G-6-PDH (E. coli):  300 µkat/L;
preservative
c. Glucose HK (GLUC3 REF 04404483).
d. Diluent NaCl 9 % (REF 04489357).
e. Calibrator (REF 10759350).
f. Sample vials and false bottom tubes.
g. Printer paper.
h. Toner cartridge.
i. Refer to MSDS located in the Operator’s Manual for
reagent description and composition
2. Other materials
a. Powder free hypoallergenic latex examination gloves.
b. Biohazardous waste storage bags and boxes (Jefferson
Smufit Corporation, Highland, IL).
c. Viro Research Envirocide Disinfectant Decontaminant
Cleaner (Fisher Scientific, St. Louis, MO)
d. Transfer pipettes (Fisher Scientific, St. Louis, MO)
e. Single fold paper towels (Ft. Howard Corp. Co, Green Bay,
WI).
f. Kim Wipe lintless tissues (Kimberly Clark Corp., Roswell,
GA).
g. Low and high in-house plasma controls (low and high,
respectively).
h. Bio-Rad controls
i. Volumetric pipettes for measuring different volumes.
iii. Equipment Maintenance
1. Roche Cobas c311– Routine maintenance
a. Perform daily maintenance as outlined in Cobas c311
manual every time machine is used.
b. Perform weekly maintenance as outlined in Cobas c311
manual once a week (generally whenever machine is used)
c. Perform monthly, quarterly and bi-annually maintenance
when assigned.
2. Record all daily, weekly, monthly, and periodical maintenance in
the “C311 Maintenance” binder. This paperwork is approved by
the supervisor monthly and filed in “C311 Maintenance” binder.
3. Roche – Preventative maintenance is performed every 6 months.
4. Pipettes are to be calibrated once a year.
5. Temperatures for the refrigerators and freezers where specimens,
controls and calibrators are stored are to be recorded. Any
readings that fall outside acceptable ranges are to be reported to
the supervisor or delegate for corrective action. These areas are
monitored by 7 day 24 hour temperature recorders for minimum
and maximum temperatures and are checked daily (working day)
by a technician to make sure the temperatures are within
acceptable ranges. The charts are to be checked and initialed by
the supervisor or delegate at least weekly.
iv. Storage Requirements – Reagent Use and Storage:

1. GLUC3
a. Shelf life at 2-8 °C: see expiration date on cobas c pack label
b. On-board, in use and refrigerated on the analyzer: 8 weeks
2. All reagents are ready for use; to load packs onto machine, follow
instructions in Cobas c311 analyzer Basic Operator Training Guide.
v. Reagent Labeling: Reagents, calibrators, controls, and solutions should be

traceably identified to indicate the following:
1. Content and quantity, concentration or titer
2. Storage requirements.
3. Date
4. The below should be followed for reagents used daily
a. Preparation date or opened and the identity of the
preparer;
b. Tech’s initials
5. When new reagents are received they must be initialed and dated
by the technician who receives them. When the reagents are
opened they are initialed and dated with open date by the
technician.
a. Reagent packs loaded onto the machine are logged by the
machine with the load date and will flag when expired.
d. Calibration:
i. Preparation and Stability
1. Material: C.f.a.s (Calibrator for automated systems) manufactured
by Roche Cobas. Human serum with chemical additives and
material of biological origin as specified on package insert.
2. Calibrator values are given in the electronically available value
sheets, organized by lot. Determinations were performed under
strictly standardized conditions on multiple Roche analyzers using
Roche system reagents and C.f.a.s. master calibrator or reference
materials.
a. Barcodes are given with each package of calibrator. They
may be used to scan the calibrator lot into the c311
3. Carefully open a bottle avoiding the loss of lyophilized material.
Using a 3 mL volumetric pipette or equivalent, add in exactly 3.0
mL of distilled/deionized water. Carefully close the bottle and
dissolve the contents completely by occasional swirling within 30
minutes. Avoid formation of foam.
4. Storage and Stability
a. Store unopened C.f.a.s at 2-8oC
i. Stability of lyophilized calibrator at 2-8oC: see
expiration date
b. Stability of glucose in the reconstituted calibrator
i. 15-25 oC: 8 hours
ii. 2-8 oC: 2 days
o
iii. (-15)—(-25 C) : 4 weeks (when frozen once)
ii. Frequency of Calibration:

1. Calibration verification is performed when a system is first placed
in service and at least every six months (or as specified by the
manufacturer) thereafter for all quantitative tests. Recalibration is
required (regardless of the length of time since last performed)
immediately if any of the following occurs;
a. A change of reagent lots for chemically or physically active
or critical components, unless the laboratory can
demonstrate that the use of different lots does not affect
the accuracy of patient/client test results or the range used
to report patient/client test data
b. If QC materials reflect an unusual trend or shift or are
outside of the laboratory's acceptable limits, and other
means of assessing and correcting unacceptable control
values fail to identify and correct the problem
c. After major maintenance or service. The Laboratory
Director must determine what constitutes major
maintenance or service.
iii. Calibration Procedure: To calibrate the c311 for glucose

1. Calibrators: S1: diH20
S2: C.f.a.s
2. Calibration Mode: Linear 2-point calibration
3. Choose the “Calibration” tab
4. Select the glucose test
5. Select “2 point” under the method window
6. Insert calibrators and controls into the correct vial positions. These
positions can be found in the Calibration load list report and QC
load list report
7. Select “Save”
8. Select the “Start” button
9. Traceability: This method has been standardized against ID/MS.
10. Calibration Acceptability Criteria
a. When QC results fail to meet the acceptable criteria, check
the sample cup containing the QC specimen for bubbles
and reanalyze the QC specimen.
b. If the QC results meet the acceptable criteria, accept the
run and report the results.
c. If steps above do not result in correction of the "out-of-
control" values for QC materials, troubleshoot the
instruments and reagents until the system is back “in
control”
d. Recalibrate the system using a new vial of glucose standard.
e. Reanalyze the calibrator, controls, and specimens.
Specimens are stable at 4-8 °C overnight. If the system
requires more than 24 hours before it can be restored to
functionality, use new aliquots of standard, controls, and
specimens for analysis.
f. Refer to Operator’s Manual for additional troubleshooting
guidance (attached in this procedure) and the Appendix
section for c311 Quick Reference guide.
g. If the calibration is still unsuccessful speak with your
supervisor for further help. Do not perform glucose analysis
until the system is declared "in-control" again.
iv. Calibration Verification – The LN2 survey is purchased through CAP.
The LN2 Calibration Verification/Linearity survey does not span the AMR
for glucose. Therefore to verify the lower end of the AMR a serial dilution
using the lowest LN2 sample is performed using a three twofold dilutions
(e.g. 1/2, 1/4, 1/8). Aliquots are assayed in duplicate and the results
regressed in EP evaluator against a Total Error of 12% (per CAP stated
evaluation report criteria for linearity assessment).
e. Quality Control:
i. Two types of quality control (QC) systems are used in this analytical
method: 1) "sample QC" and 2) "batch QC." For sample QC, 2% of
specimens are randomly selected and analyzed either within-assay or
between-assay for quality assurance purposes. If the difference between
duplicates is greater than ±10 % of the original value, the specimen is
reanalyzed. Batch QC specimens are placed in the rack at the end of the
entire run. The following criteria are for “batch QC.”
ii. Control Materials and Stability:
1. Lyphochek Assayed Chemistry Control Levels 1 and 2
a. Prepared from human serum with added chemicals,
purified biochemical material (tissue extracts of human and
animal origin), therapeutic drugs, stabilizers and
preservatives. This product is provided in lyophilized form
for increased stability.
b. Stability
i. Store unopened controls at 2-8oC
ii. Stability of lyophilized control at 2-8oC: see
expiration date
iii. Stability of Glucose in reconstituted controls
1. 2-8 oC: 7 days
2. -70 C :o up to 3 months
2. In-house controls IHL and IHH

a. Controls were prepared by collecting 450 mL (one unit) of
whole blood from two diabetic (IHH) and two non-diabetic
(IHL) subjects. The blood was collected in blood bags
without an anticoagulant. The serum was allowed to clot
for 1 hour and then separated from the red blood cells by
centrifugation in a refrigerated centrifuge (4oC) for 25 min
at 1500 g. The serum was then removed from the red
blood cells, aliquoted in 0.5-mL portions and stored at -70
°C or colder in Nalgene cryogenic vials.
b. If the stock of these controls becomes low, another batch is
prepared in time to analyze it concurrently with the current
QC materials. The new controls are used only after their
means and the ranges have been established by performing
20 characterization runs.
c. Stability
i. At -70 oC: 13 years
ii. 2-8 oC: 72 hours
iii. 20-25 oC: 24 hours
iv. See glucose validation documents for In-house
control & manufacturer control stability studies.
iii. Preparation for analysis

1. Bring frozen samples and controls to room temperature prior to
analysis.
2. Bio-Rad Controls: Using a volumetric pipet or equivalent,
reconstitute each vial with 5 mL of distilled or deionized water.
Replace the stopper and allow this product to stand for
approximately 20 minutes swirling occasionally. Before sampling,
gently swirl the vial several times to ensure homogeneity. After
each use, promptly replace the stopper and return to the
appropriate storage condition.
3. In-House Controls: One vial of each control is thawed and used in
each assay. Reconstitution is not required for these controls.
4. Controls should be discarded after daily use.
iv. Frequency for control materials: All four batch QC controls are analyzed at
the beginning and end of each assay.
v. Mean and Ranges:

1. Daily means and ranges of the controls are calculated from at least
20 inter-assay determinations. The bias ranges of the daily means
are set at ±1 SD or the 67% confidence interval (CI); the warning
limits (WL) are the ±2 SD or the 95% CI and the control limits (CL)
are the ±3 SD or the 99% CI. For the daily ranges, the bias limit is
the mean + 1 SD with warning and control limits set at the mean +2
SD and the mean + 3 SD, respectively.
a. The means and ranges for the current controls are posted
near the c311 instrument
b. Current control value assignments and limits can be found
in the Quality Control Binder located in room M771.
2. After each assay run, all control data are recorded on the Daily
Diary Log Sheet. The results of the analysis are accepted or
rejected according to the guidelines established.
3. Two types of QC charts are used in assessing the quality of an

assay.
a. The first chart plots the mean of all the replicate
determinations in a run and compares it with the
established target mean, which is the overall mean
established by the 20 or more characteristic runs. The
guideline declares a system as "out-of-control" if any of the
following events occur:
i. The mean for one control from a single run falls
outside the 99% confidence limits.
ii. The means for two controls from a single run fall
outside the 95% confidence limits.
iii. The daily means for one control from eight
successive runs (excluding the runs in which the
mean is within ±1 SD or the bias range) fall either all
above or all below the center line.
b. The second type of QC chart plots the range of the
replicates (the difference between the highest and the
lowest value of a single control within a run) and compares
it with the established target range, which is the overall
mean of daily ranges established by the 20 or more
characteristic runs. The QC guidelines for DDL declare a
system as "out-of-control" if any of the following events
occur:
i. The daily range for one control exceeds the 99%
confidence limit.
ii. The daily ranges for two controls exceed the 95%
confidence limits.
iii. The daily ranges for one control from eight
successive runs (excluding the runs in which the
mean is within1 SD or bias range) are all above the
mean line (trend rule).
c. If a run is declared out of control, investigate the system
(instrument, standards, controls etc.) to determine the
cause of the problem. Do not perform any analysis until
the problem has been resolved.
vi. Remedial Action if QC Systems Fail to Meet Acceptable Criteria

1. When QC results fail to meet the acceptable criteria, check the
sample cup containing the QC specimen for bubbles and reanalyze
the QC specimen.
2. If the QC results meet the acceptable criteria, accept the run and
report the results.
3. If steps above do not result in correction of the "out-of-control"
values for QC materials, troubleshoot the instruments and
reagents until the system is back “in control”
4. Recalibrate the system using a new vial of glucose standard.
5. Reanalyze the calibrator, controls, and specimens. Specimens are
stable at 2-8 °C overnight. If the system requires more than 24
hours before it can be restored to functionality, use new aliquots
of standard, controls, and specimens for analysis.
6. If the above steps do not correct the "out of control" condition,
consult with the supervisor for further corrective action. Do not
perform glucose analysis until the system is declared "in-control"
again.
vii. To further validate the accuracy (and for troubleshooting purposes) of
glucose measurements on the C311, Reference Material for Clinical
Chemistry Standards (ReCCS) samples are analyzed every 6 months.
f. Procedure – Stepwise:
i. Special safety precautions:
1. Gloves and lab coat are required for handling all human blood
specimens.
2. All plastic tips, sample vials, gloves, etc. that contact blood are
considered contaminated and are to be placed in an approved
waste container.
3. Work surfaces are protected by absorbent pads. The pads are
discarded into biohazardous waste container weekly or whenever
blood contamination occurs. All work surfaces are wiped down
with Envirocide weekly.
ii. Initial processing of specimens - Clinical trial specimens are checked against
the clinical trial data sheet to insure that the patient information matches.
The specimens are placed at -70oC until ready to be tested. Barcodes are
printed by the DDL that correspond with the NHANES accession numbers on
the samples. False bottom tubes are labeled with these barcodes.
iii. To run the Roche Cobas c311:

1. Before turning on the analyzer, perform the maintenance checks
as outlined on the cobas c311 analyzer Maintenance Log under the
“Check” and “Hands-on” sections.
2. All healthcare personnel shall routinely use appropriate barrier
precautions to prevent skin and mucous membrane exposure
when contact with blood or other body fluids of any patient is
anticipated. All products or objects that come in contact with
human or animal body fluids should be handled, before and after
cleaning, as if capable of transmitting infectious diseases. Wear
appropriate Personal Protective Equipment (PPE), including facial
protection, gloves, and protective clothing. Dispose of all biological
samples and diluted specimens in a biohazard waste container at
the end of analysis. Dispose of all liquid hazardous waste in
properly labeled hazardous waste container.
3. Turn on the power to the analyzer and control unit. The Analytical
Unit must be turned on before the control unit.
4. Log-in to the control unit using name: adm and password: adm
5. The system will perform the “Power On Pipe” automatically.
6. Ensure all “Push-button” maintenance functions were performed
by checking the utility screen.
7. Perform any maintenance functions that have expired
8. Put machine into maintenance mode by flipping switch on the
analyzer. Perform any necessary maintenance outlined in the
Maintenance Log that has not already been done.
9. Return machine to operation mode
10. Go to “System Overview” screen. Ensure that temperature of the
incubator is within 37oC ± 0.1oC by clicking “AU” button and
observing the core temperature.
11. Check the “Work Flow Guide” area at the top of the screen. If any
of the first five buttons is highlighted, perform that action (Daily
Maintenance, Sample Data Clear, Regent Preparing, Calibration
and QC Select, Parameter Download). These actions should be
automatically completed by the analyzer.
12. Calibrate if necessary following the calibration procedures.
13. Enter controls to run as samples
a. Go to Utility menu
b. Click on sample information
c. Type in control name
d. Hit enter
e. Select “Gluc” test button
f. Select “Save” test button
g. Enter “Barcode Read Error” mode
h. Enter name of control and position on machine
i. Repeat for all 8 controls (beginning and end)
14. Enter samples and tests to run
a. Go to Utility menu
b. Click on sample information
c. Scan barcode for sample
d. Hit enter
e. Select “Gluc” test button
f. Select “Save” test button
g. Repeat for all samples
15. To run samples, choose “Start” button at lower right of the screen
16. Hit “Start” when prompted
17. To manually shut down the instrument, choose the “Deactivate
system” option.
18. Preliminaries:
a. All reagents should be at room temperature before assay.
b. Allow frozen reference standards, QC specimens, and any
frozen blood samples to thaw. Mix all samples at least ten
minutes before putting on the machine
19. Sample preparation: Using printed labels, label the appropriate
sample vials with the corresponding sample identification, and
place the whole, uncapped original tube into the sample vials.
20. After run is completed, print hard copy of the results and proceed
to reporting results.
iv. Calculations:
1. Roche cobas c systems automatically calculate the analyte
concentration of each sample.
a. Conversion factors: mmol/L x 18.02 = mg/dL
mmol/L x 0.1802 = g/L
mg/dL x 0.0555 = mmol/L
v. Reporting results:
1. All replicate values of QC data plus all pertinent assay information
(date of analysis, reagent lot number, technician ID, samples ID
etc.) are recorded in the Microsoft Access Glucose Daily Diary Log
database located on the network drive. The calibrator value is also
recorded. Enter the data under the form “Diary Sheet Entry
Form”. The Microsoft Access program will automatically calculate
the daily mean and range for each control and determine if a run is
accepted or rejected. The current above or below the mean trend
is also calculated.
2. Any comments associated with the specimen are entered in the
comment field. If a result is below the assay detection limit, or a
sample is missing, or if the sample volume is less than 200 µL, or
the sample is otherwise unacceptable, the result field is left blank
or a –1 is entered and an appropriate comment is entered in the
assay comment field.
3. After analysis the results, date analyzed and tech initials are
imported from the instrument into the SQL server database via
secure transfer.
4. Data check sheets are printed out and checked against the
instrument printouts by the supervisor or delegate (signed by a
supervisor or delegate).
5. After results are cleared by the supervisor or delegate a result file
in the specified format is exported and uploaded to Westat via
secure transfer.
g. Reference Ranges:
i. ADA reference ranges for fasting individuals3
1. Normal: < 100 mg/dL
2. Pre-diabetic (impaired fasting glucose): 100-125 mg/dL
3. Diabetic: ≥ 126 mg/dL
ii. ADA reference ranges for 2-hour postprandial individuals3

1. Normal: < 140 mg/dL
2. Pre-diabetic (impaired glucose tolerance): 140-199 mg/dL
3. Diabetic: ≥ 200 mg/dL
h. Carryover Studies- Performed following manufacturer’s experimental design and

guidelines;
1. Prepare two specimens, one with a very high %HbA1c (≥ 16 %,
preferred HbA1c; ≥ 18.5%) and one with a very low %HbA1c value
(≤ 4.8%, preferred HbA1c; ≤ 3.8%).
2. Aliquot these specimens: 11 with low % HbA1c concentration and
10 with high %HbA1c concentration.
3. While assaying these 21 samples, other specimens or tests should
be assayed on the instrument and the samples should be assayed
in the following carryover order, otherwise the experiment is
invalid:
3 L (low) / 2 H (high)/ 1 L /2 H/4 L/2 H /1 L/2 H/1 L/2 H/1 L
Note: Ensure at least 1 set of QC is analyzed within the run to

ensure validity of results.
4. After analyzing the low and high specimens in the above order,
enter the results in EP Evaluator using the carryover program.
Refer to the EP Evaluator carryover report interpretation guide for
additional details. o, p
5. The carryover test passes if the results of the High-low sequences
are statistically identical to the results of the low-low sequences
(three times the SD of the low-low result – the SD that would be
expected if no high results were measured). The results of the low-
level sample should not be affected by the high-level sample.
6. Expected Performance:
i. No more than 0.2 % HbA1c carryover from H-L into L-L sample
with samples beyond the selected low to high % HbA1c range
is considered acceptable.
7. Carryover studies should be performed, as applicable, as part the
initial evaluation of an instrument. Carryover studies should be
repeated after major instrument maintenance (as specified by
manufacturer) or repair of the pipetting assembly.
j. Procedures for Abnormal Results:

i. Critical Value Adults and Children: < 40mg/dL or > 400mg/dL
ii. Critical results must be repeated and verified.
1. Glucose results do not have STAT turnaround. We typically receive
these samples at least 24 hours after collection. Therefore, there
is no “panic situation”.
k. Reporting Format:
i. Results are expressed on the report as mg/dL
ii. Measuring range for Plasma: 2-728 mg/dL without dilution. Glucose
results that exceed 728 mg/dL are reported as “> 728 mg/dL.”
iii. Lower detection limit: 2 mg/dL
a. The lower detection limit represents the lowest measurable
analyte level that can be distinguished from zero.
iv. Linearity data are found in the c311 Maintenance Binder under the
Linearity tab. Linearity and AMR verification studies are performed at least
twice per year refer to DDL’s QM Program for additional details.
l. Supervisor Responsibility:
i. The supervisor or delegate ensures quality control passes within the
acceptable ranges prior to releasing patient results.
m. Limitations of the Procedure/ Interfering Substances:

i. Criterion: Recovery within ± 10 % of initial value at a glucose
concentration of 70.3 mg/dL.
ii. Plasma
1. Icterus: No significant interference up to an I index of 60
(approximate conjugated and unconjugated bilirubin
concentration: 60 mg/dL.)
2. Hemolysis: No significant interference up to an H index of 1000
(approximate hemoglobin concentration: 1000 mg/dL.
3. Lipemia (Intralipid): No significant interference up to an L index of
1000. There is poor correlation between the L index (corresponds
to turbidity) and triglycerides concentration.
4. Drugs: No interference was found at therapeutic concentrations
using common drug panels.7,8
5. In very rare cases gammopathy, in particular type IgM
(Waldenström's macroglobulinemia), may cause unreliable results.
iii. Actions Required

1. Special Wash Programming: The use of special wash steps is
mandatory when certain test combinations are run together on
Roche/Hitachi cobas c systems. Refer to the latest version of the
carry over evasion list found with the NaOH/SMS/Multiclean/SCCS
Method Sheet and the operator manual for further instructions.
2. Where required a special wash or carry over evasion programming
must be implemented prior to reporting results with this test.
IV. Attachments:
a. C311 Operator’s Manual
V. References, Regulatory References, Related Documents, or Links
1. Sacks DB. Carbohydrates. In: Tietz NW, ed. Fundamentals of Clinical Chemistry.
4th ed. Philadelphia: WB Saunders 1996:351-374.
2. Knudson PE, Weinstock RS. Carbohydrates. In: Henry JB, ed. Clinical Diagnosis and
Management by Laboratory Methods. 20th ed. Philadelphia: WB Saunders
2001:211-223.
3. Sacks DB. Carbohydrates. In: Burtis CA, Ashwood ER, eds. Tietz Textbook of
Clinical Chemistry. 3rd ed. Philadelphia: WB Saunders 1999:750-785.
4. Kunst A, Draeger B, Ziegenhorn J. In: Bergmeyer. Methods of Enzymatic Analysis,
3rd ed. Volume VI, Metabolites 1: Carbohydrates. 1984:163-172.
5. Roche Diagnostic Technical support: 1-800-428-2336
VI. Appendix

65e804280dd4340bc20b10ee - Glucose SOP 6-2-22

Uploaded by

Copyright:

Available Formats

65e804280dd4340bc20b10ee - Glucose SOP 6-2-22

Uploaded by

Document Information

Original Title

Copyright

Available Formats

Share this document

Share or Embed Document

Sharing Options

Did you find this document useful?

Is this content inappropriate?

Copyright:

Available Formats

65e804280dd4340bc20b10ee - Glucose SOP 6-2-22

Uploaded by

Copyright:

Available Formats

Title: Diabetes Diagnostic Laboratory - Glucose Testing using the

Roche Cobas c311

Glucose is the major carbohydrate present in the peripheral blood. Glucose

G-6-P + NADP+ gluconate-6-P + NADPH + H+

b. Specimen Collection and Handling

iii. Specimen Stability

2. Unacceptable specimen criteria

iv. Handling Conditions

c. Equipment and Materials

iv. Storage Requirements – Reagent Use and Storage:

v. Reagent Labeling: Reagents, calibrators, controls, and solutions should be

ii. Frequency of Calibration:

iii. Calibration Procedure: To calibrate the c311 for glucose

2. In-house controls IHL and IHH

iii. Preparation for analysis

v. Mean and Ranges:

3. Two types of QC charts are used in assessing the quality of an

vi. Remedial Action if QC Systems Fail to Meet Acceptable Criteria

iii. To run the Roche Cobas c311:

ii. ADA reference ranges for 2-hour postprandial individuals3

h. Carryover Studies- Performed following manufacturer’s experimental design and

3 L (low) / 2 H (high)/ 1 L /2 H/4 L/2 H /1 L/2 H/1 L/2 H/1 L

Note: Ensure at least 1 set of QC is analyzed within the run to

j. Procedures for Abnormal Results:

m. Limitations of the Procedure/ Interfering Substances:

iii. Actions Required

V. References, Regulatory References, Related Documents, or Links

You might also like