Lp-PLA2 FS*

Order Information Reagent Preparation

Cat. No. Kit size Reagent 2 and reagent 3 must be premixed before use. Due to
1 7181 99 10 922 50 (1 x 50) hygroscopic components, reagent 3 shall be stored tightly closed,
and should not stand open for longer than 5 min. Bring reagents to
Intended Use room temperature before mixing. Make sure that there is no air
bubble on the bottom of the reagent vial R3 by tapping the vial two
Diagnostic reagent for quantitative in vitro determination of Lp-
to three times on the table.
PLA2 (lipoprotein-associated phospholipase A2) in human serum
or heparin plasma on automated DiaSys respons®910. Transfer 250 μL R3 into the R2 compartment of the twin container
by pipetting.
Summary Mix very gently to avoid foaming. In case of precipitation, leave
Lipoprotein-associated phospholipase A2 (Lp-PLA2), also known as premixed reagent until it is completely homogenized.
platelet-activating factor acetylhydrolase (PAF AH), is a calcium- Stability of premixed R2/R3: 8 weeks if stored at 2 – 8°C.
independent phospholipase released by inflammatory cells in
atherosclerotic plaques. In circulation, Lp-PLA2 is predominantly Materials Required
associated with LDL particles whereas only a small portion of
General laboratory equipment
enzyme is associated with HDL. Lp-PLA2 hydrolyzes oxidized LDL
to generate two atherogenic and inflammatory compounds:
Lysophosphatidylcholine (lyso-PC) and oxidized free fatty acids
Specimen
(oxFFA). Both substances play a major role in the development of Human serum or heparin plasma
vulnerable atherosclerotic plaques. Concentration of Lp-PLA2 is Stability [6]:
independent of the presence of other cardiovascular risk factors, 2 days at 20 – 25°C
shows minimal biovariability and is not elevated in systemic 4 weeks at 2 – 8°C
inflammatory reactions. Lp-PLA2 is a beneficial indicator for 3 months at –20°C
cardiovascular disease (CVD) risks, and may represent a potential
therapeutic target for the reduction of such risks. [1-4] Only freeze once. Discard contaminated specimens.

Method Calibrators and Controls

UV test using 1-myristoyl-2-(4-nitrophenylsuccinyl)-sn-glycero-3- DiaSys TruCal Lipid calibrator is recommended for calibration.
phosphocholine TruCal Lipid calibrator values have been made traceable to the
molar extinction coefficient of 4-nitrophenol. Use DiaSys TruLab L
Lp-PLA2 hydrolyzes the sn-position of the substrate 1-myristoyl-2-
Level 1 and Level 2 for internal quality control. Each laboratory
(4-nitrophenylsuccinyl)-sn-glycero-3-phosphocholine producing
should establish corrective action in case of deviations in control
4-nitrophenylsuccinate. After degradation in aqueous solution,
recovery.
4-nitrophenol develops which can be detected photometrically. Lp-
PLA2 activity is determined by a change in absorbance at the Cat. No. Kit size
defined wavelengths. TruCal Lipid 1 3570 99 10 045 3 x 2 mL
TruLab L Level 1 5 9020 99 10 065 3 x 3 mL
Reagents TruLab L Level 2 5 9030 99 10 065 3 x 1 mL
Components and Concentrations Note: For reconstitution of TruLab L Level 2, add exactly 1 mL of
R1: Buffer pH 7.6 < 500 mmol/L distilled water. Reconstitution of TruLab L Level 1 should be done
EDTA < 50 mmol/L according to the instruction supplied with the product.
R2: Buffer pH 2.7 < 200 mmol/L Replacement labels are attached to the reagent kit to identify
R3: Alcohol 99% TruLab L Level 2 with reduced reconstitution volume.
1-myristoyl-2-(4-nitrophenylsuccinyl)- < 200 mmol/L
sn-glycero-3-phosphocholine Performance Characteristics
Storage and Stability Exemplary data mentioned below may slightly differ in case of
deviating measurement conditions.
The reagents are stable up to the date of expiry indicated on the
kit, if stored at 2 – 8°C and contamination is avoided. Do not freeze Measuring range up to 2000 U/L.
reagent R3, protect from light and moisture. In case of higher activities re-measure samples after manual
Warnings and Precautions dilution with NaCl solution (9 g/L) or use rerun function.

1. Reagent 3: Warning. Contains: Diethylene glycol. H302 Limit of detection** 50 U/L

Harmful if swallowed. H373 May cause damage to organs Onboard stability 10 days
through prolonged or repeated exposure. P260 Do not Calibration stability 10 days
breathe mist/vapours/spray. P264 Wash hands and face
thoroughly after handling. P314 Get medical advice/attention Interfering substance Interferences Analyte
if you feel unwell. ≤ 10% up to concentration
2. In very rare cases, samples of patients with gammopathy [U/L]
might give falsified results [5]. Ascorbic acid 60 mg/dL 455
3. Please refer to the safety data sheets and take the necessary
precautions for the use of laboratory reagents. For diagnostic 60 mg/dL 907
purposes, the results should always be assessed with the Bilirubin (conjugated) 50 mg/dL 446
patient’s medical history, clinical examinations and other 50 mg/dL 884
findings.
4. For professional use only. Bilirubin (unconjugated) 50 mg/dL 414
50 mg/dL 860
Waste Management
Hemoglobin 1000 mg/dL 425
Refer to local legal requirements.
1000 mg/dL 904
N-acetylcysteine (NAC) 1500 mg/L 445
1500 mg/L 898
Lipemia (triglycerides) 1800 mg/dL 415
1800 mg/dL 932

Lp-PLA2 FS – Page 1 844 7181 10 02 74 April 2021/1

 Precision
Within run (n=20) Sample 1 Sample 2 Sample 3
Mean [U/L] 287 577 842
CV [%] 2.29 1.41 1.80
Total Precision CLSI Sample 1 Sample 2 Sample 3
(n=80)
Mean [U/L] 273 539 778
CV [%] 3.70 3.86 3.56
Method comparison (n=100)
Test x DiaSys Lp-PLA2 FS
Test y DiaSys Lp-PLA2 FS (improved)
Slope 0.958
Intercept 4.21
Coefficient of correlation 0.995
** according to CLSI document EP17-A, Vol. 24, No. 34

Conversion Factor
Lp-PLA2 [U/L] x 0.0167 = Lp-PLA2 [µkat/L]

Reference Range [6]

Adults
Men < 639 U/L
Women < 507 U/L
Each laboratory should check if the reference ranges are
transferable to its own patient population and determine own
reference ranges if necessary.

Literature
1. Ridker, P.M.; MacFadyen, J.G.; Wolfert R.L.; Koenig W.
Relationship of lipoprotein-associated phospho-lipase A2
mass and activity with incident vascular events among
primary prevention patients allocated to placebo or to statin
therapy: An analysis from the JUPITER trial. Clin Chem 2012;
58(5):877-886.
2. Münzel, T.; Gori, T. Lipoprotein-associated phospholipase A2,
a marker of vascular inflammation and systemic vulnerability.
Eur Hear J 2009; 30:2829-2831.
3. Madjid, M.; Ali, M.; Willerson, J.T. Lipoprotein-associated
phospholipase A2 as a novel risk marker for cardiovascular
disease. Tex Heart Inst J 2010; 37(1): 25-39.
4. Mannheim, D; Herrmann, J et al. Enhanced expression of Lp
PLA2 and Lysophosphatidylcholine in Symptomatic Carotid
Atherosclerotic Plaques. Stroke 2008;39:1448-1455.
5. Bakker AJ, Mücke M. Gammopathy interference in clinical
chemistry assays: Mechanism, detection and prevention. Clin
Chem Lab Med 2007; 45(9): 1240-1243.
6. Personal communication from Prof. Dr. med. Karl Winkler,
Universitaetsklinikum Freiburg, Germany.
DiaSys Diagnostic Systems GmbH
Alte Strasse 9 65558 Holzheim Germany
[Link]

* Fluid Stable

Lp-PLA2 FS – Page 2 844 7181 10 02 74 April 2021/1

Lp-PLA2 FS
Application for serum and plasma samples
This application was set up and evaluated by DiaSys. It is based on the standard equipment at that time and does not
apply to any equipment modifications undertaken by unqualified personnel.
Identification Results
This method is usable for analysis: Yes Decimals 2
Twin reaction: No Units U/L
Name: LpPLA2 Correlation factor-Offset 0.000
Shortcut: Correlation factor-Slope 1.000
Reagent barcode reference: 065
Host reference: Range
Gender Male
Technic Age
Type: Linear kinetic SERUM <=639
First reagent:[µL] 200 URINE
Blank reagent Yes PLASMA <=639
Sensitive to light CSF
Second reagent:[µL] 50 Whole blood
Blank reagent Yes Gender Female
Sensitive to light Age
Main wavelength:[nm] 405 SERUM <=507
Secondary wavelength:[nm] 508 URINE
Polychromatic factor: 1.000 PLASMA <=507
1 st reading time [min:sec] 6:00 CSF
Last reading time [min:sec] 8:12 Whole blood
Reaction way: Increasing
Linear Kinetics Contaminants
1.50
Substrate depletion: Absorbance limit Please refer to r910 Carryover Pair Table
Linearity: Maximum deviation [%]
Fixed Time Kinetics Calibrators details
Substrate depletion: Absorbance limit Calibrator list Concentration
Endpoint Cal. 1/Blank 0
Stability: Largest remaining slope Cal. 2 *
Prozone Limit [%] Cal. 3
Cal. 4
Reagents Cal. 5
Decimals Cal. 6
Units Max delta abs.
Cal. 1 0.010
Sample Cal. 2 0.005
Diluent System water Cal. 3
Hemolysis: Cal. 4
Agent [µL] 0 (no hemolysis) Cal. 5
Cleaner Cal. 6
Sample [µL] 0 Drift limit [%] 0.8

Technical limits
Calculations
Concentration technical limits-Lower 50 Model X
Concentration technical limits-Upper 2000 Degree 1
SERUM
Normal volume [µL] 2 * Enter calibrator value
Normal dilution (factor) 1
Below normal volume [µL] 10
Below normal dilution (factor) 1
Above normal volume [µL] 2
Above normal dilution (factor) 6
URIN
Normal volume [µL] 2
Normal dilution (factor) 1
Below normal volume [µL] 10
Below normal dilution (factor) 1
Above normal volume [µL] 2
Above normal dilution (factor) 6
PLASMA
Normal volume [µL] 2
Normal dilution (factor) 1
Below normal volume [µL] 10
Below normal dilution (factor) 1
Above normal volume [µL] 2
Above normal dilution (factor) 6
CSF
Normal volume [µL] 2
Normal dilution (factor) 1
Below normal volume[ µL] 10
Below normal dilution (factor) 1
Above normal volume [µL] 2
Above normal dilution (factor) 6
Whole blood
Normal volume [µL] 2
Normal dilution (factor) 1
Below normal volume[ µL] 10
Below normal dilution (factor) 1
Above normal volume [µL] 2
Above normal dilution (factor) 6

Application respons®910 April 2021/1