MAGLUMI Vitamin B12 (CLIA)

INTENDED USE
The kit has been designed for the quantitative determination of
Vitamin B12 in human serum.
The method can be used for samples over the range of
Shenzhen New Industries Lotus Medical Equipment 12.5-2000.0 pg/ml.
Biomedical Engineering Co., Ltd Limited The test has to be performed on MAGLUMI Fully-auto
No.16, Jinhui Road, 26B Cameron Court, chemiluminescence immunoassay (CLIA) analyzer (Including
Pingshan New District, Cork Street, Dublin 8, Maglumi 600, Maglumi 800, Maglumi 1000, Maglumi 1000 Plus,
Shenzhen, 518122, P.R.China. Ireland Maglumi 2000, Maglumi 2000 Plus, Maglumi 4000).
Tel: +86-755-21536601 Tel: +353-1-6571034 Catalog Number Specification
Fax:+86-755-28292740 E-mail: peter@lotusme.org 130213002M 100 tests
130613002M 50 tests

SUMMARY AND EXPLANATION OF THE TEST

Vitamin B12, also called cobalamin, is a water-soluble vitamin.
Cobalamin can be found in animal products such as meat, eggs,
FOR PROFESSIONAL USE ONLY
milk and other dairy products. During the intake process,
Store at 2-8 °C
cobalamin conjugates with a factor known as the combining
binding-protein in the stomach, then absorbed by the ileum.
CAUTION: COMPLETELY READ THE Intrinsic factor is necessary for absorption process, once getting
INSTRUCTIONS BEFORE PROCEEDING into the circulation; the cobalamin is stored in the liver. When being
required, the cobalamin will be released into the plasma by the
VB12 binding protein.
SYMBOLS EXPLANATIONS VB12 is normally involved in the metabolism of every cell of the
AUTHORISED REPRESENTATIVE IN body, especially affecting the DNA synthesis and regulation but
THE EUROPEAN COMMUNITY also fatty acid synthesis and energy production. However, many
(though not all) of the effects of functions of VB12 can be replaced
MANUFACTURER by sufficient quantities of folic acid (vitamin B9), since VB12 is
used to regenerate folate in the body. Most VB12 deficiency
symptoms are actually folate deficiency symptoms, since they
CONSULT INSTRUCTIONS FOR USE include all the effects of pernicious anemia and megaloblastosis,
which are due to poor synthesis of DNA when the body does not
have a proper supply of folic acid for the production of thymine.
KIT COMPONENTS The lack of VB12 can be caused by a variety of reasons. The most
common reason is a lack of intrinsic factor, resulting in that VB12
cannot be absorbed from food. This condition is called anemia,
IN VITRO DIAGNOSTIC MEDICAL
DEVICE mostly affecting people over the age of 50 years. The gastric
resection is another reason of VB12 deficiency, after surgery or
BATCH CODE malabsorption caused by various bacteria or inflammatory disease
affecting the small intestine. VB12 uptake has a positive
correlation with the functional length of small intestine. The lack of
CATALOGUE NUMBER
VB12 is rarely caused by abnormal dietary, unless no animal food
is consumed in years.
USE BY
PRINCIPLE OF THE TEST
TEMPERATURE LIMITATION Competitive chemiluminescence immunoassay:
( STORE AT 2-8 °C) Use ABEI to label purified VB12 antigen, use FITC to label VB12
binding-protein, use an anti-FITC polyclonal antibody to coat
magnetic microbeads. The sample (or calibrator/control, if
SUFFICIENT FOR
applicable), ABEI Label, FITC Label and magnetic microbeads
are mixed thoroughly and incubated at 37°C, forming complexes;
after precipitation in a magnetic field, decant the supernatant, and
KEEP AWAY FROM SUNLIGHT perform a wash cycle. Subsequently, the starter 1+2 is added to
initiate a flash chemiluminescent reaction. The light signal is
measured by a photomultiplier within 3 seconds as RLU which is
THIS WAY UP proportional to the concentration of VB12 present in samples.

KIT COMPONENTS
Material Supplies
Component 100 tests 50 tests
Magnetic Microbeads: coated with
sheep anti-FITC polyclonal antibody, 2.5 ml 2.0 ml
containing BSA, 0.09% NaN3.

099 Vitamin B12-IFU-V3.06-en-EU 1/4

 Calibrator Low: phosphate buffer • The prepared displacing reagent can only be used for 72 hours
containing BSA and VB12 antigen, 3.0 ml 2.0 ml at 2-8°C. After 72 hours, the displacing reagent will be expired,
0.09% NaN3. and it should be discarded
Calibrator High: phosphate buffer • Displacing reagent shelf life will be shorted if place on the
containing BSA and VB12 antigen, 3.0 ml 2.0 ml analyzer, it is recommended to finish the displacing reagent in
0.09% NaN3. 48 hours to ensure its performance.
Displacing Reagent (preparation Storage and Stability
 Sealed: Stored at 2-8° C until the expiration date.
required)
 Opened: Stable for 4 weeks. To ensure the best kit performance,
FITC Label: VB12 binding-protein
labeled with FITC, containing BSA, it is recommended to place opened kits in the refrigerator if it’s not
13.5 ml 7.5 ml
0.09% NaN3. going to be used on-board during the next 12 hours.
ABEI Label: purified VB12 antigen
labeled with ABEI, containing BSA, 13.5 ml 7.5 ml
THIS WAY UP
0.09% NaN3.
Prepare Displacing Reagent before using the integral
KEEP AWAY FROM SUNLIGHT

Reagent Vials in kit box

Empty bottle of displacing reagent 6 bottles
CALIBRATION AND TRACEABILITY
1) Traceability
Displacing Solution: 1 mmol/L NaOH. 15.0 ml
To perform an accurate calibration, we have provided the test
DTT: lyophilized 30mg DTT, reconstituted with
30.0 mg calibrators standardized against the WHO 1st International
300μl distilled water
Internal Quality Control: phosphate buffer Standard 03/178.
containing BSA and VB12 antigen, 0.09% 2) 2-Point Recalibration
2.0 ml Via the measurement of calibrators, the predefined master curve is
NaN3. (For target value, refer to Quality
Control Information date sheet) adjusted (recalibrated) to a new, instrument-specific measurement
level with each calibration
Internal quality control is only applicable with MAGLUMI system.
3) Frequency of Recalibration
For instructions for use and target value, refer to Quality Control
 After each exchange of lot (Reagent Integral or Starter
Information date sheet. User needs to judge results with their own
Reagents).
standards and knowledge.
 Every three days and/or each time a new Integral is used

(recommended).
Accessories Required But Not Provided
 After each servicing of MAGLUMI Fully-auto
MAGLUMI Reaction Module REF: 630003
chemiluminescence immunoassay (CLIA) analyzer.
MAGLUMI Starter 1+2 REF: 130299004M
 If controls are beyond the expected range.
MAGLUMI Wash Concentrate REF: 130299005M
 Whenever room temperature changes exceed 5 °C
MAGLUMI Light Check REF: 130299006M (recommended).
Please order accessories from Shenzhen New Industries
Biomedical Engineering Co., Ltd (SNIBE) or our representative. SPECIMEN COLLECTION AND PREPARATION
Sample material: serum
Collect samples using standard procedures. Store at 2-8°C: 24
Preparation of the Reagent Integral hours, for longer storage periods: freeze to below - 20°C
Avoid repeated freezing and thawing more than one time, stored
Before the sealing is removed, gentle and careful horizontal
samples should be thoroughly mixed prior to use (Vortex mixer).
shaking of the Reagent Integral is essential (avoid foam formation!)
Please ask local representative of SNIBE for more details if you
Remove the sealing and turn the small wheel of the magnetic
have any doubt.
microbeads compartment to and fro, until the color of the
Specimen Conditions
suspension has changed into brown. Place the Integral into the
• The sample which has been placed at the room temperature
reagent area and let it stand there for 30 min. During this time, the
more than 8 hours cannot be used again.
magnetic microbeads are automatically agitated and completely
• The sample serum with high concentration of protein
resuspended.
(>90g/L) cannot be used to do the tests. As the high dose of
Do not interchange integral components from different
protein will form the gel and block the needle
reagents or lots!
 •Do not use specimens with the following conditions:
Preparation of Displacing Reagent
(a) heat-inactivated specimens;
1. Use 300 μl distilled water to dissolve the lyophilized DTT
(b) Cadaver specimens or body fluids other than human
completely in DTT reagent vial, then aliquot into 3 empty tubes by
serum;
100μl each (Eppendof tube, 0.5 ml type is recommended). The
(c) Obvious microbial contamination.
dissolved DTT tube should be sealed and stored at -20°C and can
 Use caution when handling patient specimens to prevent
be stable for 2 month. Take it out to room temperature before
cross contamination. Use of disposable pipettes or pipette tips
preparing Displacing Reagent.
is recommended.
2. Pipette 5ml NaOH into the empty displacing reagent bottle, then
 Inspect all samples for bubbles. Remove bubbles with an
pipette 100μl DTT solution from DTT tube and add in the displacing
applicator stick prior to analysis. Use a new applicator stick for
reagent bottle. Using the NaOH solution in the bottle to rinse the wall,
each sample to prevent cross contamination.
make sure DTT solution is dissolved with NaOH solution completely.
 Serum specimens should be free of fibrin, red blood cells or
Then horizontally shake the bottle in round cycles for mixing,
other particulate matter.
avoided forming bubbles.
 Ensure that complete clot formation in serum specimens has
3. Place Displacing Reagent bottle into the 4th position of the kit
taken place prior to centrifugation. Some specimens,
integral. This bottle can be used for about 40 tests. (It is
especially those from patients receiving anticoagulant or
recommended customer collect enough samples and run the tests
thrombolytic therapy, may exhibit increased clotting time. If the
together).
specimen is centrifuged before a complete clotting, the
Note:
099 Vitamin B12-IFU-V3.06-en-EU 2/4
 presence of fibrin may cause erroneous results. the microbeads requires mixing to re-suspend microbeads
that have settled during shipment.
Preparation for Analysis  For microbeads mixing instructions, refer to the KIT
 Patient specimens with a cloudy or turbid appearance must be COMPONENTS, Preparation of the Reagent Integral section
centrifuged prior to testing. Following centrifugation, avoid the of this package insert.
lipid layer (if present) when pipetting the specimen into a  To avoid contamination, wear clean gloves when operating a

sample cup or secondary tube. reagent kit and sample.

 Specimens must be mixed thoroughly after thawing by low • Pay attention to the residual liquids which has dried on the kit
speed vortexing or by gently inverting, and centrifuged prior to surface.
use to remove red blood cells or particulate matter to ensure  For detailed handling precautions during system operation,

consistency in the results. Multiple freeze-thaw cycles of refer to the SNIBE service information.
specimens should be avoided.
 All samples (patient specimens or controls) should be tested TEST PROCEDURE
within 3 hours of being placed on board the MAGLUMI To ensure proper test performance, strictly adhere to the operating
System. Refer to the SNIBE service for a more detailed instructions of MAGLUMI Fully-auto chemiluminescence
discussion of onboard sample storage constraints. immunoassay (CLIA) analyzer. Each test parameter is identified
Storage via a RFID tag on the Reagent Integral. For further information
 If testing will be delayed for more than 8 hours, remove serum
please refer to the operating instructions of MAGLUMI Fully-auto
from the serum separator, red blood cells or clot. Specimens chemiluminescence immunoassay (CLIA) analyzer .
removed from the separator gel, cells or clot may be stored up 100 μl Sample , Calibrator
to 12 hours at 2-8°C. +100 μl Displacing reagent
 Specimens can be stored up to 30 days frozen at -20° C or 2 min Incubation
colder. +110 μl ABEI label
Shipping
+110 μl FITC label
 Before shipping specimens, it is recommended that +20 μl Magnetic microbeads
specimens be removed from the serum separator, red blood
15 min Incubation
cells or clot. When shipped, specimens must be packaged
400 μl Wash cycle
and labeled in compliance with applicable state, federal and
3 s Measurement
international regulations covering the transport of clinical
specimens and infectious substances. Specimens must be
shipped frozen (dry ice). DILUTION
Sample dilution by analyzer is not available in this reagent kit
WARNING AND PRECAUTIONS FOR USERS Samples with concentrations above the measuring range can be
diluted manually. After manual dilution, multiply the result by the
dilution factor.
 For use in IN-VITRO diagnostic procedures only. Please choose applicable diluents or ask SNIBE for advice before
 Package insert instructions must be carefully followed. manual dilution must be processed.
Reliability of assay results cannot be guaranteed if there are
any deviations from the instructions in this package insert. QUALITY CONTROL
 Observe quality control guidelines for medical laboratories
Safety Precautions  Use suitable controls for in-house quality control. Controls
CAUTION: This product requires the handling of human should be run at least once every 24 hours (a run cannot
specimens. exceed 24 hours), once per reagent kit and after every
 All samples, biological reagents and materials used in the
calibration. The control intervals should be adapted to each
assay must be considered potentially able to transmit laboratory’s individual requirements. Values obtained should
infectious agents. They should therefore be disposed of in fall within the defined ranges. Each laboratory should
accordance with the prevailing regulations and guidelines of establish guidelines for corrective measures to be taken if
the agencies holding jurisdiction over the laboratory, and the values fall outside the range.
regulations of each country. Disposable materials must be
incinerated; liquid waste must be decontaminated with sodium
hypochlorite at a final concentration of 5% for at least half an
LIMITATIONS OF THE PROCEDURE
hour. Any materials to be reused must be autoclaved using an 1) Limitations
overkill approach. A minimum of one hour at 121°C is usually Assay results should be utilized in conjunction with other clinical and
considered adequate, though the users must check the laboratory data to assist the clinical decision-making for individual
effectiveness of their decontamination cycle by initially patients.
validating it and routinely using biological indicators. A skillful operation and strict adherence to the instructions are
 It is recommended that all human sourced materials be
necessary to obtain reliable results.
considered potentially infectious and handled in accordance Procedural directions must be followed exactly and careful operation
with the 29 CFR. 1910.1030 Occupational exposure to must be used to obtain valid results. Any modification of the
bloodborne pathogens. Biosafety Level 2 or other appropriate procedure is likely to alter the results.
biosafety practices should be used for materials that contain Bacterial contamination or repeated freeze-thaw cycles may affect
or are suspected of containing infectious agents. the test results.
 This product contains Sodium Azide; this material and its
2) Interfering Substances
container must be disposed of in a safe way. The assay is unaffected by bilirubin<20 mg/dl, haemoglobin<150
 Safety data sheets are available on request.
mg/dl or triglycerides< 3000 mg/dl.
3) HAMA
Handling Precautions Patient samples containing human anti-mouse antibodies (HAMA)
 Do not use reagent kits beyond the expiration date.
may give falsely elevated or decreased values. Although
 Do not mix reagents from different reagent kits.
HAMA-neutralizing agents are added, extremely high HAMA
 Prior to loading the Reagent Kit on the system for the first time,
serum concentrations may occasionally influence results.

099 Vitamin B12-IFU-V3.06-en-EU 3/4

 E 1200
RESULTS F 2000
1) Calculation of Results
 The analyzer automatically calculates the FA concentration in 6) Method comparison
each sample by means of a calibration curve which is A comparison of MAGLUMI VB12 (y) with a commercially
generated by a 2-point calibration master curve procedure. available VB12 test (x) using clinical samples gave the following
The results are expressed in pg/ml. For further information correlations (pg/ml):
please refer to the operating instructions of MAGLUMI Linear regression
Fully-auto chemiluminescence immunoassay (CLIA) analyzer. y = 1.092x+3.863
2) Interpretation of Results r = 0.9991
 Based on the 95% confidence interval, the reference value Number of samples measured: 100
range is: 200-1100 pg/ml The sample concentrations were between 39.61 and 1388.27
VB12 deficiency <200 pg/ml pg/ml.
 Results may differ between laboratories due to variations in

population and test method. If necessary, each laboratory REFERENCES

should establish its own reference range. 1 Pagana, Kathleen D. & Pagana, Timothy J. Mosbys Diagnostic
and Laboratory Test Reference 8th Edition: Mosby, Inc., Saint
PERFORMANCE CHARACTERISTICS Louis, MO. Pp 460-461, 999-1000, 834-836.
1) Precision 2 Clarke, W. and Dufour, D. R., Editors . Contemporary Practice
Intra-assay coefficient of variation was evaluated on 3 different in Clinical Chemistry: AACC Press, Washington, DC. Pp
levels of controls. Repeatedly measure 10 times in the same run to 407-408.
calculate the coefficient of variation. 3 Wu, A. Tietz Clinical Guide to Laboratory Tests, 4th Edition:
Intra-assay precision Saunders Elsevier, St. Louis, MO. Pp 634-635.
Control Mean(pg/ml) SD(pg/ml) CV% 4 Henry's Clinical Diagnosis and Management by Laboratory
Level 1 238.86 16.88 7.07 Methods. 21st ed. McPherson RA and Pincus MR, eds.
Level 2 621.22 47.02 7.57
Philadelphia: 2007, Pg. 506-507.
Level 3 1103.06 85.27 7.73
5 Tietz Textbook of Clinical Chemistry and Molecular Diagnostics.
Inter-assay coefficient of variation was evaluated on three batches
Burtis CA, Ashwood ER, Bruns DE, eds. St. Louis: Elsevier
of kits. Repeatedly measured 3 different levels of controls 10 times
Saunders; 2006, Pp 1100-1103.
in the same run, and 30 times for each levels to calculate the
6 Thomas, Clayton L., Editor (1997). Taber's Cyclopedic Medical
coefficient of variation.
Dictionary. F.A. Davis Company, Philadelphia, PA [18th
Inter-assay precision
Edition].
Control Mean(pg/ml) SD(pg/ml) CV%
Level 1 210.26 16.32 7.76 7 Wu, A. (2006). Tietz Clinical Guide to Laboratory Tests, Fourth
Level 2 647.13 52.29 8.08 Edition. Saunders Elsevier, St. Louis, Missouri.410-413,
Level 3 1125.68 95.46 8.48 1124-1127.
2) Analytical Sensitivity
<12.5 pg/ml.
The detection limit represents the lowest analyte level that can be
distinguished from zero.

3) Specificity
The specificity of the VB12 assay system was assessed by
measuring the apparent response of the assay to various
potentially cross reactive analytes.
Compound Concentration Cross reactivity
FA 100 ng/ml 0.2%

4) Recovery
Consider Calibrator High of known concentration as a sample,
dilute it by 1:2 ratio with diluents, and measure the diluted
concentration for 10 times. Then calculate the expected
concentration and recovery of measured concentration. The
recovery should be within 90% -110%.

Expected Mean Measuring Recovery

630.597 pg/ml 636.903 pg/ml 101%

5) Linearity
Use VB12 calibrator to prepare the six-point standard curve,
measuring all points’ RLU except point A, and then do
four-parameter linear fitting in log-log coordinate, the absolute
linear correlation coefficient(r) should be bigger than 0.9800.
Calibrator Concentration Absolute linear
Point pg/ml correlation coefficient (r)
A 0
B 150 r=0.9910
C 300
D 600

099 Vitamin B12-IFU-V3.06-en-EU 4/4