PG II iFlash

Immunoassay Analyzer

Anti-Pepsinogen II acridinium-ester-labeled
REF C86041 2×50 Tests R2 conjugate; 4.0 mL/pack; 0.05% ProClin
300.

INTENDED USE Assay diluent, 4.0 mL/pack; 0.05% ProClin

R3
The iFlash-PG II assay is a paramagnetic particle 300.
chemiluminescent immunoassay (CLIA) for the
quantitative determination of Pepsinogen II in human Calibrator 1, 1 bottle, 1.0 mL, phosphate
serum and plasma using the iFlash Immunoassay CAL1 buffer with protein stabilizers, 0.05%
Analyzer. ProClin 300.
SUMMARY AND EXPLANATION
Calibrator 2, 1 bottle, 1.0 mL, Pepsinogen II
Pepsinogen II is produced by chief cells and mucous neck CAL2 in phosphate buffer with protein stabilizers,
cells of the gastric mucosa, in pyloric glands in the gastric 0.05% ProClin 300.
antrum, and Brunner’s glands in the proximal duodenum.
The ratio of concentration of Pepsinogen I (PG I) to PG II Calibrator 3, 1 bottle, 1.0 mL, Pepsinogen II
in plasma or serum of normal subjects is about 4:1. CAL3 in phosphate buffer with protein stabilizers,
0.05% ProClin 300.
PGs are released into the circulation and it is widely
accepted that serum PG level reflects the functional and
morphologic status of stomach mucosa. Human MATERIALS REQUIRED (BUT NOT PROVIDED)
pepsinogens have a diagnostic value for various REF C89999/C89959/C89949, iFlash Pre-Trigger
gastroduodenal disorders, especially for peptic ulcer, Solution: hydrogen peroxide solution.
atrophic gastritis and gastric cancer. The Pepsinogen I/ II REF C89998/ C89958/ C89948, iFlash Trigger Solution:
ratio can provide even better information on the extent of sodium hydroxide solution.
chronic gastritis. REF C89997, iFlash Wash Buffer: phosphate buffered
ASSAY PRINCIPLE saline solution with 0.05% ProClin 300.

The iFlash-PG II assay is a sandwich immunoassay. REF C80001, iFlash Wash Buffer (10×): phosphate
buffered saline solution with 0.05% ProClin 300.

st
1 incubation: Pepsinogen II in the sample,
anti-Pepsinogen II coated paramagnetic microparticles REF C89996, reaction vessels.
react to form a complex. Controls: Commercial controls could be used.
 Wash: The unbound materials are washed away from WARNINGS AND PRECAUTIONS
the solid phase in a magnetic field.
IVD For in vitro diagnostic use

nd
2 incubation: Acridinium-ester-labeled anti-human
 No known test method can offer the complete
Pepsinogen II antibody conjugate is added to form a
assurance that products derived from human sources
sandwich complex.
will not transmit infection. Therefore, all humanized
 Another Wash. materials should be considered potentially infectious.
 Trigger of signal: The Pre-Trigger and Trigger  Exercise the normal precautions required for handling
Solutions are added to the reaction mixture. The all laboratory reagents.
resulting chemiluminescent reaction is measured as
 Disposal of all waste material should be in accordance
relative light units (RLUs).
with local guidelines.
 A direct relationship exists between the amount of
 Wear gloves when handling specimens or reagents.
anti-Pepsinogen II in the sample and the RLUs
detected by the iFlash optical system.  Clean and disinfect all spills of specimens or reagents
using a suitable disinfectant.
 Results are determined via a calibration curve, which
is instrument-specifically generated by 3-point  iFlash Trigger solution contains sodium hydroxide
calibration and a master curve provided via the (NaOH) and should be avoided contact with eyes.
reagent QR code.
REAGENT HANDLING
REAGENTS  The reagents may not be used after the stated
Reagent kit, 100 tests, 2 packs, 50 tests/pack expiration date.
Anti-Pepsinogen II coated microparticles,  Avoid the formation of foam with all reagents.
R1
3.5 mL/pack, 0.05% ProClin 300.  The reagents in the pack and calibrators are ready for
use.
 Close the bottles of calibrator right after calibration and

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PG II iFlash
Immunoassay Analyzer

store at 2–8°C. ASSAY PROCEDURE

 Do not pool reagents within a reagent kit or between  Refer to the system operating instruction or the online
reagent kits. help system for detailed information on preparing the
 Prior to loading the iFlash-PG II reagent pack on the system.
system for the first time, suspend the microparticles by  The test-specific parameters stored in barcode on the
inverting the reagent pack 30 times. reagent pack are read in. In case the barcode cannot
 For further information on reagent handling be read, enter the sequence numbers.
precautions during system operation, refer to the  Carry out calibration, if necessary.
iFlash system operating instruction.  Place the calibrators CAL1, CAL2 and CAL3 in the
calibrator rack in the sample zone. Only keep
STORAGE AND STABILITY
calibrators open during calibration.
Storage:  Test application.
 Store at 2–8°C in an upright position.  Load samples (Use 20 μL of sample for each
 The kit may be used immediately after removal from determination in addition to the sample container and
2-8°C storage. system dead volumes).
 Click RUN, the iFlash System performs all the
Stability:
functions automatically and calculates the results.
 Unopened at 2–8°C: up to the stated expiration date.
 Opened at 2–8°C: 28 days. CALIBRATION

 Store on-board: 28 days.  Traceability: This assay is traceable to commercial

reference serum.
SPECIMEN COLLECTION AND PREPARATION  Every iFlash-PG II reagent kit has a QR code label
 Serum or plasma (lithium heparin, sodium heparin containing the specific information for calibration of the
potassium EDTA, and sodium citrate) are the particular reagent lot.
recommended samples. Other anticoagulants have  To perform an iFlash-PG II calibration, test CAL1,
not been validated for use with the iFlash-PG II assay. CAL2 and CAL3 in duplicate, and the predefined
 Ensure that serum specimens to form complete clot master curve is adapted to the analyzer.
prior to centrifugation.  Once an iFlash-PG II calibration is accepted and
 Centrifuge the specimens. stored, all subsequent samples may be tested without
 Store specimens at room temperature (20 to 25°C) for further calibration unless:
no longer than 8 hours.  After 28 days when using the same reagent lot.
 If the testing will not be completed within 8 hours,  A reagent kit with a new lot number is used.
refrigerate the samples at 2 to 8°C.  Controls are out of range.
 If the testing will not be completed within 3 days, or for  Required by pertinent regulations.
shipment of samples, freeze at -20°C or colder.
 Frozen specimens must be mixed thoroughly after MEASURING RANGE
thawing.  0.50 – 150 ng/mL
 The samples may be frozen for maximum 3 times. QUALITY CONTROL
 Centrifuge specimens with a lipid layer on the top, and Quality control materials should be run as single
transfer only the clarified specimen without the lipemic determinations at least once every 24 hours when the test
material. is in use, once per reagent kit and after every calibration.
 Ensure that residual fibrin and cellular matter have Include commercially available quality control materials
been removed prior to analysis. that cover at least two levels of analyte. Follow
 Use with caution in handling patient specimens to manufacturer’s instructions for reconstitution and storage.
prevent cross-contamination. Each laboratory should establish mean values and
 Do not use heat-inactivated samples. acceptable ranges to assure proper performance. Quality
control results that do not fall within acceptable ranges
 Ensure that the patient samples, calibrators and
may indicate invalid test results.
controls are at ambient temperature (20–25°C) before
measurement. RESULT
 Due to the possible evaporation, specimens and
Calculation:
calibrators on the analyzers should be measured
within 2 hours. The iFlash system automatically calculates the analyte
concentration of each sample. The results are given in
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PG II iFlash
Immunoassay Analyzer

ng/mL. Pepsinogen II reagents, samples and controls.

The within run precision was determined by testing each
Expected Values:
sample in replicates of 10 (n = 10), and calculating percent
A study of with iFlash-PG II assay on samples from 365 coefficient of variation (%CV). The results of the study are
apparently healthy people yielded the following result: shown below:
th th
<15 ng/mL, Pepsinogen I/Pepsinogen II > 3 (2.5 -95 Sample Mean (ng/mL) SD %CV
percentile)
It is recommended that each laboratory establish its own 1 4.82 0.19 3.98
expected reference range for the specific population.
2 14.50 0.58 3.97
LIMITATIONS
3 49.94 2.28 4.57
 The iFlash-PG II assay is limited to the determination
of Pepsinogen II in human serum or plasma (lithium The between run precision was determined by testing
heparin, sodium heparin, potassium EDTA, and each sample in duplicate, two separate runs daily for 20
sodium citrate). It has not been validated for use with days (n = 80), and calculating percent coefficient of
other types of plasma. variation (%CV). The results of the study are shown
 The use of serum separator (gel) blood collection below:
tubes has been validated for use with this assay. Sample Mean (ng/mL) SD %CV
However, it is not possible to survey all manufacturers
or tube types. 1 4.99 0.23 4.61
 The upper limit of the measuring range of this assay is
2 49.14 2.58 5.24
150 ng/mL. Over-range samples may be diluted with
wash buffer recommended 1:5. Such as diluted by
hand, the result should be multiplied by the dilution Analytical Sensitivity
factor. If the machine is automatic dilution, the The detection limit representing the lowest measurable
machine will automatically calculate the results. The analyte level is 0.50 ng/mL, which can be distinguished
maximum dilution ratio of not less than 1:10. from zero. It is calculated as the value lying two standard
 If the results are inconsistent with clinical evidence, deviations above that of the lowest standard of the master
additional testing is suggested to confirm the result. curve (standard 1 + 2 SD, n = 20).
 For diagnostic purposes, the results should be Analytical Specificity
interpreted in light of the total clinical presentation of
The analytical specificity of iFlash-PG II assay was
the patient, including symptoms, clinical history results.
evaluated with analog into the negative sample.
 Specimens from heparinized patients may be partially
Analog Acceptable
coagulated and erroneous results could occur due to Analog Assay result
concentration range
the presence of fibrin. name (ng/mL)
(ng/mL) (ng/mL)
 The results from an alternative assays (i.e. EIA or RIA)
may not be equivalent and cannot be used Pepsinogen
200 < 0.5 0.03
interchangeably. Ⅰ

 The assay is unaffected by icterus (bilirubin < 30

mg/dL), hemolysis (Hb < 1,500 mg/dL), lipemia Method Comparison
(Intralipid < 1,500 mg/dL) and total serum protein (< 10 A comparison of the iFlash-PG II assay (y) with a
g/dL). commercially available Pepsinogen II assay (x) using
 No interference was observed from rheumatoid factors clinical samples was performed, and the curve is fitted
up to a concentration of 2,000 IU/mL. with Linear regression
 No interference was observed from anti-nuclear y = 1.007x + 6.165
antibodies up to a concentration of 500 U/mL. r = 0.989
 No interference was observed from HAMA up to a Sample concentration: 0.59 – 146.12 ng/mL
concentration of 600 ng/mL. Number of samples measured: 90
PERFORMANCE CHARACTERISTICS REFERENCES
Below are the representative performance data, and the 1. Kitahara F, Kobayashi K, Sato T, et al. Accuracy of
results obtained in individual laboratories may differ. screening for gastric cancer using serum pepsinogen
Precision concentrations. Gut. 1999; 44: 693-697.
The precision of iFlash-PG II was determined using 2. HHS Publication, 4th ed., May 1999. Biosafety in

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PG II iFlash
Immunoassay Analyzer

Microbiological and Biomedical Laboratories. ANNEX A:

Available. Explanation of abbreviation
3. Samloff IM, Varis K, Ihamaki T, et al. Relationships
Abbreviation Explanation
Among Pepsinogen I, Serum Pepsinogen II, and
Gastric Mucosal Histology. Gastroeneterology 1982;
83: 204-209. Product No.
4. Kikuchi S, Wade O, Miki K, et al.Serum Pepsinogens
as a Screening test of extensive chronic gastritis. Calibrator
Gastroenterologica Jpaonica.1987; 22 (2): 133-141.
5. Miki k, Oka H:Significance of determination of serum Reagent
pepsinogen. Naika 1986;58 (3): 716-718.
6. Oka H, Miki K, et al. Medical application of pepsinogen
Number of tests
RIA Kit. Rinho Seijinbyo 1989; 19 (4) 531-537.
7. Fujinaga M, Chernaia MM, Tarasova NI, Mosimann
SC, James MN. Crystal structure of human pepsin and Manufactured by
its complex with pepstatin. Protein Sci. 1995-05, 4 (5):
960–72. EU Representative

SHENZHEN YHLO BIOTECH CO., LTD. EC Declaration of Conformity

1st-4th Floor, No.5 Building, Lishan Industrial

Area, Xinghai Road, Nanshan District, Shenzhen Caution
518054, China
Instructions for use

Wellkang Ltd (www.CE-marking.eu) In vitro diagnostic medical

Suite B, 29 Harley St., London W1G 9QR, UK device

Lot No.

Date of manufacture

Expiry date

Biohazard Symbol

Pictograms for Caution

Pictograms for Hazardous to the

aquatic environment

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