AFP iFlash

Immunoassay Analyzer

REAGENTS
REF C86003 2×50 Tests Reagent kit, 100 tests, 2 packs, 50 tests/pack
Anti-AFP coated microparticles, 3.5 mL/pack,
R1
0.05% ProClin 300.
INTENDED USE
The iFlash-AFP assay is a paramagnetic particle Anti-AFP acridinium-ester-labeled conjugate; 4.0
R2
mL/pack, 0.05% ProClin 300.
chemiluminescent immunoassay (CLIA) for the
quantitative determination of Alpha1-fetoprotein (AFP) in Calibrator 1, 1 bottle, 1.0 mL, Tris buffer with
human serum and plasma using the iFlash Immunoassay CAL1
protein stabilizers, 0.05% ProClin 300.
Analyzer.
Do not use it to evaluate the risk of trisomy 21 (Down Calibrator 2, 1 bottle, 1.0 mL, AFP in Tris buffer
CAL2
syndrome). with protein stabilizers, 0.05% ProClin 300.

SUMMARY AND EXPLANATION Calibrator 3, 1 bottle, 1.0 mL, AFP in Tris buffer
CAL3
with protein stabilizers, 0.05% ProClin 300.
Alpha-fetoprotein (AFP) is an albumin-like glycoprotein
with a molecular weight of 69,000 daltons, and is formed
in the yolk sac, non-differentiated liver cells, and the fetal MATERIALS REQUIRED (BUT NOT PROVIDED)
gastro-intestinal tract. In normal condition, AFP stems REF C89999/C89959/C89949, iFlash Pre-Trigger
from the hepatic cell of embryo, and disappears from the Solution: hydrogen peroxide solution.
blood after birth in two weeks. REF C89998/ C89958/ C89948, iFlash Trigger Solution:
More than 70% of patients with primary hepatocellular sodium hydroxide solution.
carcinoma have been reported to have elevated levels of REF C89997, iFlash Wash Buffer: phosphate buffered
serum AFP. Elevated AFP levels have occasionally been saline solution with 0.05% ProClin 300.
found in association with gastrointestinal tract cancers
REF C80001, iFlash Wash Buffer (10×): phosphate
with and without liver metastases and only rarely in other
buffered saline solution with 0.05% ProClin 300.
tumors. Serum AFP has been found to be elevated during
pregnancy, in diseases such as ataxia telangiectasia, REF C89996, reaction vessels.
hereditary tyrosinemia, teratocarcinoma and in benign REF C89901, Sample Diluent A.
hepatic conditions, such as acute viral hepatitis, chronic Controls: Commercial controls could be used.
active hepatitis and cirrhosis. Elevation of serum AFP in
benign hepatic diseases is usually transient. WARNINGS AND PRECAUTIONS
IVD For in vitro diagnostic use
ASSAY PRINCIPLE
 No known test method can offer the complete
The iFlash-AFP assay is a sandwich immunoassay. assurance that products derived from human sources
 Incubation: AFP in the sample, anti-AFP coated will not transmit infection. Therefore, all humanized
paramagnetic microparticles and anti-AFP materials should be considered potentially infectious.
acridinium-ester-labeled conjugate react to form a  Exercise the normal precautions required for handling
sandwich complex. all laboratory reagents.
 Wash: The unbound materials are washed away from  Disposal of all waste material should be in accordance
the solid phase in a magnetic field. with local guidelines.
 Trigger of signal: The Pre-Trigger and Trigger  Wear gloves when handling specimens or reagents.
Solutions are added to the reaction mixture. The
 Clean and disinfect all spills of specimens or reagents
resulting chemiluminescent reaction is measured as
using a suitable disinfectant.
relative light units (RLUs).
 iFlash Trigger solution contains sodium hydroxide
 A direct relationship exists between the amount of AFP
(NaOH) and should be avoided contact with eyes.
in the sample and the RLUs detected by the iFlash
optical system. REAGENT HANDLING
 Results are determined via a calibration curve, which  The reagents may not be used after the stated
is instrument-specifically generated by 3-point expiration date.
calibration and a master curve provided via the  Avoid the formation of foam with all reagents.
reagent QR code.
 The reagents in the pack and calibrators are ready for
use.
 Close the bottles of calibrator right after calibration and
store at 2–8°C.
 Do not pool reagents within a reagent kit or between
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AFP iFlash
Immunoassay Analyzer

reagent kits. system.

 Prior to loading the iFlash-AFP reagent pack on the  The test-specific parameters stored in barcode on the
system for the first time, resuspend the microparticles reagent pack are read in. In case the barcode cannot
by inverting the reagent pack 30 times slightly. be read, enter the sequence numbers.
 For further information on reagent handling  Carry out calibration, if necessary.
precautions during system operation, refer to the  Place the calibrators CAL1, CAL2 and CAL3 in the
iFlash system operating instruction. calibrator rack in the sample zone. Only keep
calibrators open during calibration.
STORAGE AND STABILITY
 Test application.
Storage:
 Load samples (Use 20 μL of sample for each
 Store at 2–8°C in an upright position.
determination in addition to the sample container and
 The kit may be used immediately after removal from system dead volumes).
2-8°C storage.
 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.
CALIBRATION
 Opened at 2–8°C: 28 days.
 Traceability: This assay has been standardized
 Store on-board: 28 days. st
against the 1 IRP WHO Reference Standard 72/225.
SPECIMEN COLLECTION AND PREPARATION  Every iFlash-AFP 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-AFP calibration, test CAL1. CAL2,
not been validated for use with the iFlash-AFP 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-AFP 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.
MEASURING RANGE
 Frozen specimens must be mixed thoroughly after
thawing.  1.0 – 2,000 ng/mL
 The samples may be frozen for maximum 1 time. 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
acceptable ranges to assure proper performance. Quality
 Do not use heat-inactivated samples.
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 The iFlash system automatically calculates the analyte
2 hours. concentration of each sample. The results are given in
ng/mL.
ASSAY PROCEDURE
 Refer to the system operating instruction or the online Expected Values:
help system for detailed information on preparing the A study of iFlash-AFP assay on samples from 350
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AFP iFlash
Immunoassay Analyzer

apparently healthy patients of various age groups yielded The within run precision was determined by testing each
the following result: sample in replicates of 10 (n = 10), and calculating percent
th coefficient of variation (%CV). The results of the study are
< 8 ng/mL (95 percentile)
shown below:
It is recommended that each laboratory establish its own
expected reference range for the specific population. Sample Mean (ng/mL) SD %CV

LIMITATIONS 1 10.37 0.43 4.15

 The iFlash-AFP assay is limited to the determination of 2 101.94 4.82 4.73
AFP in human serum or plasma (lithium heparin,
sodium heparin, potassium EDTA, and sodium citrate). The between run precision was determined by testing
It has not been validated for use with other types of each sample in duplicate, two separate runs daily for 20
plasma. days (n = 80), and calculating percent coefficient of
 The use of serum separator (gel) blood collection tubes variation (%CV). The results of the study are shown
has been validated for use with this assay. However, it below:
is not possible to survey all manufacturers or tube Sample Mean (ng/mL) SD %CV
types.
 The upper limit of the measuring range of this assay is 1 10.05 0.43 4.28
2,000 ng/mL. Over-range samples may be diluted with
2 103.60 4.48 4.32
with Sample Diluent A at 1:50. If using manual dilution,
the results should be multiplied by the dilution multiple.
If the machine automatically dilutes, machine will Analytical Sensitivity
automatically calculate results. The detection limit representing the lowest measurable
 If the results are inconsistent with clinical evidence, analyte level is 1.0 ng/mL, which can be distinguished
additional testing is suggested to confirm the result. from zero. It is calculated as the value lying two standard
deviations above that of the lowest standard of the master
 For diagnostic purposes, the results should be
curve (standard 1 + 2 SD, n = 20).
interpreted in light of the total clinical presentation of the
patient, including symptoms, clinical history results. Method comparison
 Specimens from heparinized patients may be partially A comparison of the iFlash-AFP assay (y) with a
coagulated and erroneous results could occur due to commercially available AFP assay (x) using clinical
the presence of fibrin. samples was performed, and the curve is fitted with Linear
 The results from an alternative assays (i.e. EIA or RIA) regression
may not be equivalent and cannot be used y = 1.0329x -2.639
interchangeably.
r = 0.993
 Samples containing an apparent AFP level as high as
Sample concentration: 2.01 – 1,900 ng/mL
3,000,000 ng/mL did not exhibit a hook effect in the
Number of samples measured: 90
iFlash-AFP assay.
 The assay is unaffected by icterus (bilirubin < 30 REFERENCES
mg/dL), hemolysis (Hb < 1500 mg/dL), lipemia 1. Ruoslathi E, Engvall E, Kessler MJ. Chemical
(Intralipid < 520 mg/dL) and total serum protein (< 10 Properties of Alpha-Fetoprotein. In: Herberman RB.
g/dL).
McIntire KR (eds). Immunodiagnosis of Cancer. New
 No interference was observed from rheumatoid factors York: Marcel Dekker Inc 1979: 101–117.
up to a concentration of 2,000 IU/mL.
2. Brewer JA, Tank ES. Yolk sac tumors and
 No interference was observed from anti-nuclear alpha-fetoprotein in first year of life. Urology. 1993; 42,
antibodies up to a concentration of 500 U/mL. 1: 79–80.
 No interference was observed from HAMA up to a 3. Sato Y, et al. Early recognition of hepatocellular
concentration of 600 ng/mL. carcinoma based on altered profiles of
alphafetoprotein. New Engl J Med. 1993; 328, 25:
PERFORMANCE CHARACTERISTICS
1802–1806.
Below are the representative performance data, and the
4. Klepp O. Serum tumor markers in testicular and
results obtained in individual laboratories may differ.
extragonadal germ cell malignancies. Scand J Clin
Precision Lab Invest Suppl. 1991; 206: 28–41.
The precision of iFlash-AFP was determined using AFP 5. Fateh-Moghadam A, Stieber P. Sensible use of tumor
reagents and controls. Two controls, consisting low, and markers. Boehringer Mannheim 1994; Cat. No.
median concentration of AFP were assayed. 1536869.
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AFP iFlash
Immunoassay Analyzer

SHENZHEN YHLO BIOTECH CO., LTD.

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

Area, Xinghai Road, Nanshan District,
Shenzhen 518054, P.R. China

Wellkang Ltd (www.CE-marking.eu)

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

ANNEX A:
Explanation of abbreviation

Abbreviation Explanation

Product No.

Calibrator

Reagent

Number of tests

Manufactured by

EU Representative

EC Declaration of Conformity

Caution

Instructions for use

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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