Please only refer to the current product lot insert enclosed with the kits package for execution

and reporting

0123 130206015M: 100 tests

130606015M: 050 tests

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MAGLUMI Lp-PLA2 (CLIA)
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INTENDED USE
The kit is an in vitro chemiluminescent immunoassay for the quantitative determination of lipoprotein-associated phospholipase
A2 (Lp-PLA2) in human serum using the MAGLUMI series Fully-auto chemiluminescence immunoassay analyzer (including
Maglumi 600, Maglumi 800, Maglumi 1000, Maglumi 2000, Maglumi 2000 Plus, Maglumi 4000 and Maglumi 4000 Plus).
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SUMMARY AND EXPLANATION OF THE TEST

Lp-PLA2 (Lipoprotein-associated phospholipase A2) - also known as platelet-activating factor acetylhydrolase - is a
vascular-specific inflammatory enzyme, predominantly expressed by macrophages, lymphocytes and foam cells in
1,2
atherosclerotic plaques . Circulating Lp-PLA2 is mainly associated with apolipoprotein B-containing lipoproteins, hence closely
3,4
associated with low-density lipoprotein (LDL) . The enzyme hydrolyzes oxidized phospholipids on LDL particles within the
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arterial intima, generating two highly inflammatory mediators, lysophosphatidylcholine and oxidized non-esterified fatty acids,
5-7
with pro-inflammatory and pro-atherosclerosis effect . Therefore, the release of Lp-PLA2 can also be interpreted as an excellent
8,9
indicator of the pro-inflammatory response . By detecting the level of Lp-PLA2 in the circulatory system can be an independent
10
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predictor of cardiovascular disease .

Many important studies confirm a strong association between Lp-PLA2 levels and cardiovascular risk among different
11,12
populations . Due to the fact that Lp-PLA2 is involved in the causal pathway of plaque inflammation and plaque rupture, the
testing for Lp-PLA2 represents a valuable adjunctive tool which goes beyond traditional cardiovascular risk assessment.
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PRINCIPLE OF THE TEST

The Lp-PLA2 assay is a sandwich chemiluminescence immunoassay.
The sample (or calibrator/control, if applicable), buffer, anti-Lp-PLA2 monoclonal antibody-labeled ABEI and magnetic
microbeads coated with another anti-Lp-PLA2 monoclonal antibody are mixed thoroughly and incubated at 37°C,. forming
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sandwich of immuno-complexes. After precipitation in a magnetic field, the supernatant is decanted and then perform a wash
cycle. Subsequently, the Starter 1+2 are added to initiate a chemiluminescent reaction. The light signal is measured by a
photomultiplier within 3 seconds as relative light units (RLUs), which is proportional to the concentration of Lp-PLA2 present in
samples(or calibrator/control, if applicable).

KIT COMPONENTS
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Material provided
100 tests 50 tests
Component Contents
(REF: 130206015M) (REF: 130606015M)
Coated with anti-Lp-PLA2 monoclonal
Magnetic Microbeads 2.5 mL 2.0 mL
antibody, containing BSA, NaN3(<0.1%).
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Calibrator Low Lp-PLA2, containing BSA, NaN3 (<0.1%). 2.5 mL 2.0 mL

Calibrator High Lp-PLA2, containing BSA, NaN3 (<0.1%). 2.5 mL 2.0 mL

Buffer Containing BSA, NaN3 (<0.1%). 12.5 mL 7.5 mL

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Anti-Lp-PLA2 monoclonal antibody labeled

ABEI Label 7.5 mL 4.5 mL
with ABEI, containing BSA, NaN3 (<0.1%).
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Control 1 Lp-PLA2, containing BSA, NaN3 (<0.1%). 2.0 mL 2.0 mL

Control 2 Lp-PLA2, containing BSA, NaN3 (<0.1%). 2.0 mL 2.0 mL

All reagents are provided ready-to-use.

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Accessories Required But Not Provided

MAGLUMI Series:
Reaction Module REF: 630003
Starter 1+2 REF: 130299004M
Wash Concentrate REF: 130299005M
Light Check REF: 130299006M
Please order accessories from Shenzhen New Industries Biomedical Engineering Co., Ltd (SNIBE) or our authorized
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representatives.

CALIBRATION
Traceability: This method has been standardized against the SNIBE internal reference substance.
Test of assay specific calibrators allows the RLU values to adjust the assigned master curve. Results are determined via a
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calibration curve which is instrument-specifically generated by 2-point calibration (10 calibrations) and a master curve provided
via the reagent Radio Frequency Identification (RFID) CHIP.
Recalibration is recommended if any of the following conditions occurs:
 After each exchange of lots (Reagent or Starter Reagents).

 Every week and/or each time a new reagent kit is used (recommended).

 After instrument service is required.

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 If controls lie outside the expected range.

 Whenever room temperature changes exceed 5° C (recommended).

QUALITY CONTROL
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Follow government regulations or accreditation requirements for quality control frequency.

Internal quality control is only applicable with MAGLUMI system. For instructions for use and target value refer to Lp-PLA2
(CLIA) Quality Control Information. User needs to judge results with their own standards and knowledge.
For detailed information about entering quality control values, refer to the operating instructions of MAGLUMI series Fully-auto
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chemiluminescence immunoassay analyzer.

To monitor system performance and chart trends, commercially available quality control materials are required. Treat all quality
control samples the same as patient samples. A satisfactory level of performance is achieved when analyte values obtained are
within the acceptable Control Range for the system or within your range, as determined by an appropriate internal laboratory
quality control scheme. If the quality control results do not fall within the Expected Values or within the laboratory’s established
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values, do not report results. Take the following actions:

 Verify that the materials are not expired.

 Verify that required maintenance was performed.

 Verify that the assay was performed according to the instructions for use.

 Rerun the assay with fresh quality control samples.

 If necessary, contact your local technical support provider or distributor for assistance.
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SPECIMEN COLLECTION AND PREPARATION

 Use standard sampling tubes or tubes containing separating gel. Collect blood aseptically followed the universal precautions for
venipuncture.
 To ensure consistency in results, specimens must be transferred to a centrifuge tube and centrifuged at ≥10,000 RCF (Relative
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Centrifugal Force) for 15 minutes.

 Ensure that complete clot formation in serum specimens has taken place prior to centrifugation. Some specimens, especially
those from patients receiving anticoagulant or thrombolytic therapy, may exhibit increased clotting time.
 If the specimen is centrifuged before a complete clotting, the presence of fibrin may cause erroneous results. Samples must be
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free of fibrin and other particulate matter.

 Do not use hemolyzed or grossly lipemic specimens as well as specimens containing particulate matter or exhibiting obvious
microbial contamination. Inspect all specimens for bubbles, and remove bubbles before analysis for optimal results.
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 Avoid repeating freeze-thaw cycles. The sample can be frozen and thawed for only once. Specimens must be mixed thoroughly
after thawing.
 Centrifuged specimens with a lipid layer on the top must be transferred to a sample cup or secondary tube. Care should be taken
to transfer only the clarified specimen without the lipaemic material.
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 All samples (patient specimens and controls) should be tested within 3 hours when placed on board the MAGLUMI System.
Refer to the SNIBE service for more details discussion of onboard sample storage constraints.
 If testing will be delayed for more than 8 hours, remove serum from the serum separator, red blood cells or clot. Specimens
removed from the separator, cells or clot may be stored up to 72 hours at 2-8°C.

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 Specimens can be stored up to 60 days frozen at -20°C or colder. Stored samples should be thoroughly mixed prior to use
(Vortex mixer).
 Before shipping specimens, it is recommended that specimens be removed from the serum separator, red blood cells or clot.
When shipped, specimens should be packaged and labeled in compliance with applicable state, federal and international
regulations covering the transport of clinical specimens and infectious substances. Specimens should be shipped frozen.

WARNING AND PRECAUTIONS FOR USERS

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
 For In Vitro Diagnostic Use.
 Follow the package insert carefully. Reliability of assay results cannot be guaranteed if there are any deviations from the
instructions in this package insert.
Safety Precautions
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 CAUTION: This product requires the handling of human specimens. It is recommended that all human sourced materials be

considered potentially infectious and handled in accordance with the 29 CFR 1910.1030 Occupational exposure to bloodborne
pathogens. Biosafety Level 2 or other appropriate biosafety practices should be used for materials that contain or are
suspected of containing infectious agents.
 All samples, biological reagents and materials used in the assay should be considered potentially able to transmit infectious

agents. They should therefore be disposed of in accordance with the practices of your institution. Discard all materials in a safe
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and acceptable manner and in compliance with prevailing regulatory requirements.

 This product contains Sodium Azide. Dispose of contents and containers must be in accordance with all local, regional and

national regulations.
 Refer to safety data sheets which are available on request.
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Handling Precautions
 Do not use reagent kits beyond the expiration date.

 Do not interchange reagent components from different reagents or lots.

 Prior to loading the Reagent Kit on the system for the first time, the Reagent Kit requires mixing to re-suspend magnetic
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microbeads that have settled during shipment.

 For magnetic microbeads mixing instructions, refer to the Preparation of the Reagent section of this package insert.

 To avoid contamination, wear clean gloves when operating with a reagent kit and sample.

 Over time, residual liquids may dry on the septum surface. These are typically dried salts which have no effect on assay efficacy.

For detailed discussion of handling precautions during system operation, refer to the SNIBE service information.
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STORAGE AND STABILITY

 Sealed: Stored at 2-8°C until the expiration date.
 Opened at 2-8°C: Minimum stability is 4 weeks.
 On-board: Minimum stability is 4 weeks.
 To ensure the best kit performance, it is recommended to place opened kits in the refrigerator after the end of the intraday test
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work. It is still possible to keep on using the kit beyond the opened or on-board period if the controls are found within the
expected ranges.
 Keep upright for storage to facilitate later proper resuspension of magnetic microbeads.
 Keep away from sunlight.

TEST PROCEDURE
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Preparation of the Reagent

 Resuspension of the magnetic microbeads takes place automatically when the kit is loaded successfully, ensuring the magnetic
microbeads are totally resuspended homogenous prior to use.
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 To ensure proper test performance, strictly adhere to the operating instructions of MAGLUMI Fully-auto chemiluminescence

immunoassay analyzer. Each test parameter is identified via a RFID CHIP on the Reagent. For further information please refer
to the operating instructions of MAGLUMI series Fully-auto chemiluminescence immunoassay analyzer.
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DILUTION
Sample dilution by analyzer is not available in this reagent kit.
Samples with concentrations above the measuring range can be diluted manually. The recommended dilution is 1:9. After manual
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dilution, multiply the result by the dilution factor. Please choose applicable diluents or ask SNIBE for advice before manual dilution.
High-Dose Hook
For the Lp-PLA2 assay, no high dose hook effect was observed when samples containing Lp-PLA2 up to 10,000 ng/mL.

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 Please only refer to the current product lot insert enclosed with the kits package for execution and reporting

LIMITATION
 A skillful technique and strict adherence to the instructions are necessary to obtain reliable results.
 Bacterial contamination or heat inactivation of the specimens may affect the test results.
 A result within the expected range does not rule out the presence of disease and should be interpreted together with the
patient’s clinical picture and other diagnostic procedures.
 Test results are reported quantitatively. However, diagnosis of a disease should not be based on the result of a single test, but
should be determined in conjunction with clinical findings in association with medical judgment.
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 Any therapeutical decision should also be taken on a case-by-case basis.
 Patient samples containing human anti-mouse antibodies (HAMA) may give falsely elevated or decreased values. Although
HAMA-neutralizing agents are added, extremely high HAMA serum concentrations may occasionally influence results.

RESULTS
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Calculation of Results
The analyzer automatically calculates the Lp-PLA2 concentration in each sample by means of a calibration curve which is generated
by a 2-point calibration master curve procedure. The results are expressed in ng/mL. For further information please refer to the
operating instructions of MAGLUMI series Fully-auto chemiluminescence immunoassay analyzer.
Interpretation of Results
The expected range for the Lp-PLA2 was obtained 256 apparently healthy individuals in China, and gave the following reference
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value:
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<250 ng/mL(95 percentile).
Since there is no international standard material for Lp-PLA2, different IVD manufacturer have different traceability chain.
Therefore results from assays of other manufacturers cannot be used interchangeably. Results may differ between laboratories
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due to variations in population and test method. If necessary, each laboratory should establish its own reference range.

PERFORMANCE CHARACTERISTICS
Precision
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Precision for the Lp-PLA2 assay was determined as described in the CLSI EP5-A2. 3 human serum pools and 2 controls
containing different concentration of analyte were assayed in duplicate at two independent runs per day for 20 testing days. The
result is summarized in the following table:
Mean(ng/mL) Within-Run Between-Run Total
Sample
(N=80) SD(ng/mL) %CV SD(ng/mL) %CV SD(ng/mL) %CV
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Serum Pool 1 17.917 1.046 5.84 0.811 4.53 1.324 7.39

Serum Pool 2 279.796 10.130 3.62 9.836 3.52 14.119 5.05
Serum Pool 3 559.874 8.841 1.58 16.203 2.89 18.458 3.30
Control 1 252.046 12.986 5.15 0.000 0.00 12.986 5.15
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Control 2 469.512 14.059 2.99 13.915 2.96 19.780 4.21

Limit of Blank (LoB)

The LoB for the Lp-PLA2 assay is 1.0 ng/mL.
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Limit of Detection (LoD)

The LoD for the Lp-PLA2 assay is 2.0 ng/mL.

Measuring Range
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1.0-1000 ng/mL (defined by the limit of blank and the maximum of the master curve). Values below the limit of blank are reported
as <1.0 ng/mL. Values above the measuring range are reported as >1000 ng/mL.
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Linearity
The assay is linear between 2.0 ng/mL and 1000 ng/mL based on a study performed with guidance from CLSI EP6-A. Nine
equally distributed levels of samples were prepared by spiking a serum sample containing Lp-PLA2 1050 ng/mL with a serum
sample free of Lp-PLA2 (0.0 ng/mL).The mean sample recovery ranged between 90% to 110%.
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Method Comparison
A total of 1139 samples in the range of 11.34 to 993 ng/mL were tested by the Lp-PLA2 assay and a commercially available
2
immunoassay. The data from the resulting linear regressions are summarized as: y=0.9809x+5.3336, r =0.9887.

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Analytical Specificity
Studies were performed to compare serum containing the following compounds at the indicated concentrations
with serum samples. The table below lists the substances tested and the concentration at which no significant interference was
observed:
Cross-Reactant Concentration
Atorvastatin 140 μmol/L
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Clopidogrel sulfate 140 μmol/L
Aspirin 3300 μmol/L
Fenofibrate 125 μmol/L
Pravastatin 10 μmol/L
Vitamin C 227 μmol/L
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Lisinopril 0.74 μmol/L

Endogenous Interference
Substances up to the following concentrations did not interfere with the assay:
 Bilirubin 40 mg/dL
 Triglyceride 1000 mg/dL
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 Hemoglobin 2000 mg/dL

 HSA 70 mg/mL
 RF 1500 IU/mL
 HAMA 30 ng/mL
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 ANA +++ (High positive sample)

REFERENCES
1. Hakkinen T, Luoma JS, et al. (1999). Arterioscler Thromb Vasc Biol 19: 2909-17.
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2. Kudo I and Murakami M. (2002). Prostaglandins Other Lipid Mediat 68-69: 3-58.
3. Caslake MJ, Packard CJ, et al. (2000). Atherosclerosis 150: 413-9.
4. Witztum JL. (1994). Lancet 344: 793-5.
5. Chisolm GM and Steinberg D. (2000). Free Radical Biol Med 28: 1815-26.
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6. Kolodgie FD, Burke AP, et al. (2006). Arterioscler Thromb Vasc Biol 26: 2523-9.
7. Macphee CH and Suckling KE. (2002). Expert Opin Ther Targets 6: 309-14.
8. Macphee CH. (2001). Curr Opin Pharmacol 1: 121-5.
9. Macphee CH, Moores KE, et al. (1999). Biochem J 338: 479-87.
10. Packard CJ, O’Reilly DS, et al. (2000). N Engl J Med 343: 1148-55. 309.
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11. Lerman A and McConnell JP (2008). Am J Cardiol 101 (Suppl): 11F-22F.

12. Wolfert RL, Kim NW, et al. (2004). Circulation 110: Suppl 3:

Shenzhen New Industries Biomedical Engineering Co., Ltd.

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No.16, Jinhui Road, Pingshan New District, Shenzhen, 518122, P.R. China
Tel: 0086-755-21536601 Fax: 0086-755-28292740
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Lotus Global Co., Ltd.

1 Four Seasons Terrace, West Drayton, Middlesex London, UB7 9GG, United Kingdom
Tel: 0044-20-75868010 Fax: 0044-20-79006187
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SYMBOLS EXPLANATIONS

Consult instructions for use Manufacturer

Temperature limit
Use-by date
( Store at 2-8 °C)
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Contains sufficient for Keep away from sunlight

Authorized representative in the

This way up
European Community
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In vitro diagnostic medical device Kit components

Catalogue number Batch code

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