PG I iFlash

Immunoassay Analyzer

Assay diluent, 4.0 mL/pack; 0.05% ProClin

REF C86040 2×50 Tests R3
300.

Calibrator 1, 1 bottle, 1.0 mL, phosphate

INTENDED USE
CAL1 buffer with protein stabilizers, 0.05% ProClin
The iFlash-PG I assay is a paramagnetic particle
chemiluminescent immunoassay (CLIA) for the 300.
quantitative determination of Pepsinogen I in human
serum and plasma using the iFlash Immunoassay Calibrator 2, 1 bottle, 1.0 mL, Pepsinogen I
CAL2 in phosphate buffer with protein stabilizers,
Analyzer.
0.05% ProClin 300.
SUMMARY AND EXPLANATION
Calibrator 3, 1 bottle, 1.0 mL, Pepsinogen I
Pepsinogens are inactive precursors of pepsins which are CAL3 in phosphate buffer with protein stabilizers,
proteolytic enzymes in gastric juice, and are 0.05% ProClin 300.
immunologically classified into Pepsinogen I (PG I) &
Pepsinogen II (PG II). PG I is produced by the fundic
MATERIALS REQUIRED (BUT NOT PROVIDED)
gland, and PG II is produced by the fundic gland, the
REF C89999/C89959/C89949, iFlash Pre-Trigger
cardiac glands, pyloric glands and Brunner’s glands.
Solution: hydrogen peroxide solution.
PG I level reliably correlates with the number of chief cells
REF C89998/ C89958/ C89948, iFlash Trigger Solution:
in the gastric corpus mucosa. Correspondingly, the loss of
sodium hydroxide solution.
chief cells results in a linear decrease in PG I. The loss of
chief cells is, on the other hand, a result of atrophic REF C89997, iFlash Wash Buffer: phosphate buffered
gastritis. So, PG I testing method is used for gastric mass saline solution with 0.05% ProClin 300.
screening in physical examination. REF C80001, iFlash Wash Buffer (10×): phosphate
buffered saline solution with 0.05% ProClin 300.
ASSAY PRINCIPLE
REF C89996, reaction vessels.
The iFlash-PG I assay is a sandwich immunoassay.
Controls: Commercial controls could be used.

st
1 incubation: Pepsinogen I in the sample,
anti-Pepsinogen I coated paramagnetic microparticles WARNINGS AND PRECAUTIONS
react to form a complex. IVD For in vitro diagnostic use
 Wash: The unbound materials are washed away from  However, as no known test method can offer the
the solid phase in a magnetic field. complete assurance that products derived from human

nd
2 incubation: Acridinium-ester-labeled anti-human source will not transmit infection. Therefore, all
Pepsinogen I antibody conjugate is added to form a humanized materials should be considered potentially
sandwich complex. infectious.
 Another Wash.  Exercise the normal precautions required for handling
 Trigger of signal: The Pre-Trigger and Trigger all laboratory reagents.
Solutions are added to the reaction mixture. The  Disposal of all waste material should be in accordance
resulting chemiluminescent reaction is measured as with local guidelines.
relative light units (RLUs).  Wear gloves when handling specimens or reagents.
 A direct relationship exists between the amount of  Clean and disinfect all spills of specimens or reagents
anti-Pepsinogen I in the sample and the RLUs using a suitable disinfectant.
detected by the iFlash optical system.
 iFlash Trigger solution contains sodium hydroxide
 Results are determined via a calibration curve, which (NaOH) and should be avoided contact with eyes.
is instrument-specifically generated by 3-point
calibration and a master curve provided via the REAGENT HANDLING
reagent QR code.  The reagents may not be used after the stated
expiration date.
REAGENTS
 Avoid the formation of foam with all reagents.
Reagent kit, 100 tests, 2 packs, 50 tests/pack
 The reagents in the pack and calibrators are ready for
Anti-Pepsinogen I coated microparticles, 3.5
R1 use.
mL/pack, 0.05% ProClin 300.
 Close the bottles of calibrator right after calibration and
Anti-Pepsinogen I acridinium-ester-labeled store at 2–8°C.
R2
conjugate; 4.0 mL/pack; 0.05% ProClin 300.
 Do not pool reagents within a reagent kit or between
reagent kits.
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PG I iFlash
Immunoassay Analyzer

 Prior to loading the iFlash-PG I 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
STORAGE AND STABILITY calibrator rack in the sample zone. Only keep
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).

Stability:  Click RUN, the iFlash System performs all the

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 a commercial
available kit.
SPECIMEN COLLECTION AND PREPARATION
 Every iFlash-PG I 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 I calibration, test CAL1, CAL2
not been validated for use with the iFlash-PG I assay.
and CAL3 in duplicate, and the predefined master
 Ensure that serum specimens to form complete clot curve is adapted to the analyzer.
prior to centrifugation.
 Once an iFlash-PG I calibration is accepted and stored,
 Centrifuge the specimens. all subsequent samples may be tested without further
 Store specimens at room temperature (20 to 25°C) for 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.  1.0 – 250 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
 Ensure the patient samples, calibrators and controls control results that do not fall within acceptable ranges
are at ambient temperature (20–25°C) before may indicate invalid test results.
measurement.
RESULT
 Due to the possible evaporation, specimens and
calibrators on the analyzers should be measured Calculation:
within 2 hours. The iFlash system automatically calculates the analyte
concentration of each sample. The results are given in
ASSAY PROCEDURE ng/mL.
 Refer to the system operating instruction or the online
help system for detailed information on preparing the

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PG I iFlash
Immunoassay Analyzer

Expected Values: sample in replicates of 10 (n = 10), and calculating percent

A study of with iFlash-PG I assay on samples from 300 coefficient of variation (%CV). The results of the study are
apparently healthy people yielded the following result: shown below:
67-200 ng/mL, Pepsinogen I /Pepsinogen II > 3 Sample Mean (ng/mL) SD %CV
th th
(2.5 -95 percentile)
1 11.03 0.49 4.44
It is recommended that each laboratory establish its own
expected reference range for the specific population. 2 105.73 3.23 3.05

LIMITATIONS 3 199.64 8.87 4.44

 The iFlash-PG I assay is limited to the determination of
Pepsinogen I 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
below:
 The use of serum separator (gel) blood collection
tubes has been validated for use with this assay. Sample Mean (ng/mL) SD %CV
However, it is not possible to survey all manufacturers
1 10.58 0.55 5.16
or tube types.
 The upper limit of the measuring range of this assay is 2 104.95 6.97 6.64
250 ng/mL. Over-range samples may be diluted with
wash buffer recommended 1:5. Such as diluted by Analytical Sensitivity
hand, the result should be multiplied by the dilution
The detection limit representing the lowest measurable
factor. The maximum dilution ratio of not less than
analyte level is 1.0 ng/mL, which can be distinguished
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
interpreted in light of the total clinical presentation of Analytical Specificity
the patient, including symptoms, clinical history results. The analytical specificity of iFlash-PG I assay was
 Specimens from heparinized patients may be partially evaluated with analog into the negative sample.
coagulated and erroneous results could occur due to Analog Acceptable Assay
the presence of fibrin. Analog name concentratio range result
n (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
interchangeably. Pepsinogen II 100 < 1.0 0.76
 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 I assay (y) with a
g/dL).
commercially available Pepsinogen I 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 = 1.012x + 8.118
antibodies up to a concentration of 500 U/mL.
r = 0.991
 No interference was observed from HAMA up to a
Sample concentration: 1.72 – 230.24 ng/mL
concentration of 600 ng/mL.
Number of samples measured: 90
PERFORMANCE CHARACTERISTICS
REFERENCES
Below are the representative performance data, and the
results obtained in individual laboratories may differ. 1. Kitahara F, Kobayashi K, Sato T, et al. Accuracy of
screening for gastric cancer using serum pepsinogen
Precision concentrations. Gut. 1999; 44: 693-697.
The precision of iFlash-PG I was determined using 2. Fujinaga M, Chernaia MM, Tarasova NI, Mosimann
Pepsinogen I reagents, samples and controls. SC, James MN. Crystal structure of human pepsin and
The within run precision was determined by testing each its complex with pepstatin. Protein Sci. 1995-05, 4 (5):

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PG I iFlash
Immunoassay Analyzer

960–72. ANNEX A:
3. Steinberg W. The clinical utility of the CA 19-9 Explanation of abbreviation
tumor-associated antigen. Am J Gastroenterol. 1990;
Abbreviation Explanation
85 (4): 350–355.
4. HHS Publication, 4th ed., May 1999. Biosafety in
Microbiological and Biomedical Laboratories. Product No.
Available .
5. Samloff IM, Varis K, Ihamaki T, et al. Relationships Calibrator
Among Pepsinogen I, Serum Pepsinogen II, and
Gastric Mucosal Histology. Gastroeneterology 1982; Reagent
83 :204-209.
6. Miki K, Ichinose M, Shimizu A,et,al. Serum
Number of tests
Pepsinogens as a Screening test of Extense chronic
gastritis. Gastroenterologica Japonica. 1987; 22 (2):
133-141. Manufactured by

SHENZHEN YHLO BIOTECH CO., LTD. EU Representative

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

Area, Xinghai Road, Nanshan District, EC Declaration of Conformity
Shenzhen 518054, P.R. China
Caution

Wellkang Ltd (www.CE-marking.eu)

Instructions for use
Suite B, 29 Harley St., London W1G 9QR, UK
In vitro diagnostic medical
device

Lot No.

Date of manufacture

Expiry date

Biohazard Symbol

Pictograms for Caution

Pictograms for Hazardous to the

aquatic environment

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