BODITECH MED INC. I-CHROMA AFP-25

BODITECH MED INC. i-CHROMA AFP-25

BodiTech Med’s express and implied warranties (including impliedwarranties of merchantability and fitness) are conditional uponobservance of BodiTech Med’s published directions with respect to the useof BodiTech Med’s products.

For Technical Assistance call

Boditech Med’s Technical Services at

Tel: +82 (33) 243-1400

E-mail:

BodiTech Med Incorporated

G-Tech Village, 1144-2 Geoduri Dongnaemyeon,

Chuncheon, Gangwondo200-883

Republic of Korea

Tel: +82 (33) 243-1400

Fax: +82 (33) 243-9373

EU Representative:Jai Jun Choung, Ph.D.

EU Biotech Development Ltd.

81 Oxford Street, LONDON, W1D 2EU
United Kingdom
Tel: +44 207 903 5441

Fax: +44 207 903 5333

E-Mail:

i-CHROMATM is registered trademarks of BodiTech MedIncorporated.

Revision No: 05

Date of last revision: May 13, 2010.

BodiTech Med Inc.

i-CHROMATM AFP

ImmunoAssay for Quantitative Measurement of Alpha Feto Protein(AFP) in Human Whole blood with i-CHROMATM Reader System.

INTENDED USE

The i-CHROMATM AFP Test along with i-CHROMATM Reader is a fluorescence immunoassay that measures AFP in human whole blood.

INTRODUCTION

Alpha-fetoprotein (AFP) is a 1-globulin family of human plasma proteins and a glycoprotein with a molecular weight approximately 70 kDa. AFP is produced primarily in the liver of developing fetus. It can be found in maternal blood and in amniotic fluid since it is secreted into fetal serum. The concentration of AFP in healthy adult is below 15 ng/ml but it shows a great increase in several malignant diseases, mostly primary hepatocelluar carcinoma and non-seminomatous testicular cancer. Some 70-90% of patients with primary hepatocelluar carcinoma and nonseminomatous testicular cancer have been observed to have high levels of serum AFP. High levels of serum AFP also have been found in a limited number of patients diagnosed with various diseases such as gastrointestinal tract cancer, viral hepatitis, chronic active hepatitis, alcoholic cirrhosis, and adenocarcinomas of lung, pancreas, and gall bladder. Since AFP is well known to be an important prognostic indicator of non-seminomatous testicular cancer, its most definitive role is on monitoring post-treatment clinical status and the post therapeutic evaluation of patients.

PRINCIPLE

The i-CHROMATM AFP Testis based on fluorescence immunoassay technology. The i-CHROMATM AFP Test uses a sandwich immunodetection method, such that by mixing detector buffer with blood specimen in test vial, the fluorescence-labeled detector anti-AFP antibodyin buffer binds to AFP antigen in blood specimen. As the sample mixture is loaded onto the sample well of the test device and migrates the nitrocellulose matrix of test strip by capillary action, the complexes of detector antibody and AFP are captured to anti-AFP sandwich pair antibody that has been immobilized on test strip. Thus the more AFPantigen is in blood specimen, the more complexes are accumulated on test strip. Signal intensity of fluorescence of detector antibody reflects amount of AFP captured and is microprocessed from i-CHROMATM Reader to show AFP concentration in blood specimen. The default result unit of i-CHROMATM AFP Testis displayed as an ng/L from i-CHROMATM Reader. The working range and the detection limit of i-CHROMATM AFP Test system are 5 - 350 ng/mL and 5ng/mL respectively.

* Reference Value : 20 ng/mL

COMPOSITION OF REAGENTS

Thei-CHROMATM AFPTestKit consists of Test Device and Detector Buffer. Test Device is individually sealed with a desiccant in aluminum pouch, and Detector Buffer is packed and delivered separately from Test Device in a styrofoam box filled with ice pack.

•Test Device contains a test strip in which murine monoclonal antibody against human AFP andrabbit IgG have been immobilized on the test and on the control line of strip, respectively.

• Detector Buffer contains fluorescence-labeled anti-AFP (Mouse monoclonal), fluorescence-labeled anti-rabbit IgG, BSA as a stabilizer, and Sodium Azide as a preservative in PBS.

WARNINGS AND PRECAUTIONS

• IVD For In Vitro Diagnostic Use.

•Carefully follow the instructions and procedures described in this insert. REF Catalog No.i-CHROMATM AFP-25

• Don’t use Test Device if its lot # does not match with ID Chip # that is inserted onto the instrument.

• Thei-CHROMATM AFP TestKit is only operational in thei-CHROMATM Reader. Andtests should be applied by professionally trainedstaff working in certifiedlaboratories at some remove from the patient and clinic at which the sample(s) is taken byqualified medical personnel.

•LOTNeither inter-change materials from different product lots nor use beyond the expiration date. The use of medical device beyond expiration date may affect on test result.

•Thei-CHROMATM AFP TestDevice should remain in itsoriginal sealed pouch until ready to use. Do notuse the Test Device if the pouch is damaged or the seal is broken. Discardafter single use.

•AFP Test Deviceand Reader should be used away from vibration and magnetic field. During normal usage, i-CHROMATM AFP Test may introduce minute vibration, which should be regarded normal.

•Use separate clean pipette tipsand sample vials fordifferent specimens.The pipette tips and sample vials should be used for one specimen only. Discardafter single use.

•Blood specimens, used test devices, pipette tips and sample vials arepotentially infectious. Proper laboratory safety techniques, handling and disposalmethods shouldbe followed in accordance with standard procedures and relevant regulations observed by microbiological hazard materials.

•The test will be applied on a routine basis and not in emergencysituations.

• Do not smoke, eat, or drink in areas in which specimens or kit reagents are handled.

STROAGE AND STABILITY

•Store the detector buffer in a refrigerator at 2° - 8°C. The Detector Buffer is stable up to 20 months if stored in a refrigerator.

•Once removed from refrigerator, allow the Detector Buffer for 30 minutes to return to room temperature before testing.

• Storei-CHROMATM AFP TestDevice at 4°-30°C in its sealed pouch. The i-CHROMATM AFP TestDeviceis stable for 20 months (while in the sealed pouch) ifstored at 4°-30°C.

•If stored in a refrigerator, allow a minimum of 30 minutesfor the Test Device to reach room temperature while it is in the sealedpouch.

•Do not remove the device from the pouch until ready to use.The Test Device should be used immediately once opened.

• The storage and shipping of Test Kit should be complied as indicated in manual. However, it is remotely possible that only part of Test Kit is affected bystability problems.

SAMPLE COLLECTIONANDPREPARATION

The test can be performed with either serum or plasma or whole blood.

•For serum sample, collect the blood in a tube without anticoagulant and allow to be clotted. Remove the serum from the clot as soon as possible to avoid hemolysis. For plasma sample, collect the blood in a tube treated with EDTA. Anticoagulants other than EDTA for plasma specimen have not been evaluated. If testing cannot be conducted within an hour after preparation of specimen, the serum/plasma should be stored at -20o C until tested. In case of use whole blood, apply it immediately after specimen was taken.

•The specimen must be at room temperature and be homogeneous before testing. Frozen specimens must be completely thawed, thoroughly mixed, and brought to room temperature prior to testing. If specimens are to be shipped, they should be packed in compliance with regulations.

•It is recommended to avoid using severely hemolyzed specimens whenever possible. If a specimen appears to be severely hemolyzed, another specimen should be obtained and tested.

MATERIALS PROVIDED

BodiTech Med Incorporated i-CHROMATM AFP Test

REF Catalog No. i-CHROMATMAFP -25

Kit contains:

Test Devices 25T/box

Detector Buffer 1vial (2ml/vial)

ID Chip 1/box

Insert 1 ea

MATERIALSREQUIRED BUT NOT PROVIDED

i-CHROMATM Reader REF Catalog No.FR-203

Thermal Printer

Transfer pipette (10, 20, 75µL size)

PROCEDURE

• Image of the test kit

window sample well

1. Set a Test Device on a dust-free clean place.
2. Check/insert ID Chip onto the instrument. Make sure that the Test Device lot # matches with ID Chip #.
3. Take out one tube of DetectionBufferfrom refrigerator and leave it at room temperature.
4. Draw 30µL of whole blood(15µLof serum, plasma or Control) with a transfer pipette and add it to the tube containing Detector Buffer.
5. Mix well the specimen with Detector Buffer by tapping or inverting the tube.
6. Take 75 µL of sample mixture and load it onto the well of disposable Test Device.
7. Leave the Test Device at room temperature for 15 min before inserting the device into the holder.
8. To start scanning, insert test device onto the holder ofi-CHROMATM Reader and press “SELECT” button.. Make sure direction of Test Device and push the device back all the way. The instrument will automatically start to scan the Test Device immediately.
9. Read the results on the display screen of i-CHROMATM Reader.

Refer to i-CHROMATMReaderOperation Manual for the complete instructions on use of the Test.

RESULT

The i-CHROMATM Reader calculates AFPtest results automatically and displays AFP concentration on the screen as form of mg/L.For further information, refer to the Operation Manual for the i-CHROMATM Reader.

Quality Control

Quality Control

A quality control test using commercially available controls should be performed as a part of good testing practice, to confirm the expected QC results, to confirm the validity of the assay, and to assure the accuracy of patient results. If you want to perform QC of Test Kit, we recommend using Abbott Axsym control.

A quality control test should be performed at regular intervals, and before using a new kit with patient specimens, controls should be tested to confirm the test procedure, and to verify the tests produce the expected QC results. QC specimens should also be run whenever there is any question concerning the validity of results obtained. Upon confirmation of the expected results, the test device is ready to use with patient specimens. Control standards are not provided with this test kit. For information about obtaining the controls, contact BodiTech Med Incorporate’s Technical Services for assistance.

Procedure Control

Each i-CHROMATM AFP TestDevice contains internal control thatsatisfies routine quality control requirements. Thisinternal control is performed each time a patient sample is tested. This controlindicates that the test device was inserted and read properly by i-CHROMATM Reader.An invalid result from the internal control causes anerror message on i-CHROMATM Reader indicating that the test should berepeated.

LIMITATIONS OF THE PROCEDURE

•The results of i-CHROMATM AFP Test should be evaluated with all clinicaland laboratory data available. If AFP Test results do not agree with the clinical evaluation, additional tests should be performed.

• The false positive results include cross-reactions withsome components of serum from individual to antibodies; and non-specific adhesionof some components in human blood that have similar epitopes to capture and detector antibodies. In the case of false negativeresults, the most common factors are: non-responsiveness of antigen to the antibodies by that certain unknown components are masking its epitope, such that antigen cannot be seen by the antibodies; instability of AFP antigen,resulting in degradation with time and, or temperature, such that they become no longerrecognizable by antibodies; anddegraded other test components. The effectiveness of thetestis highlydependent on storage of kits and sample specimens at optimal conditions.

• Plasma using anticoagulants (e.g. heparin or citrate) other than EDTA has not been evaluated in i-CHROMATM AFP Testand thus should not be used.

•Other factors may interfere with i-CHROMA TM AFP Test and may causeerroneous results. These include technical or procedural errors, as well asadditional substances in blood specimens.

PERFORMANCE CHARACTERISTICS

1. Analytical Sensitivity: Analytical sensitivity means the lowest concentration of AFP that the test system can detect with CV<10%. Analytical sensitivity of i-CHROMATM AFP Test was determined by testing 10 times with three lots of reagents. Analytical sensitivity of i-CHROMATM AFP Test system was 5 ng/mL.

2. Specificity:Other bio-molecules, such as Hb, CEA, PSA, ALT, Troponin I, CK-MB, Albumin, and serum amyloid P component were added to test specimen with much higher level than their physiological level in normal blood.

3. Imprecision: For the intra-assay imprecision, 20 replicates were tested at each control sample. For the inter-assay imprecision, tests were conducted on 10 sequential days, with 10 runs per day and with 10 replicates at each AFP concentration.

Imprecision of i-CHROMATM AFPTest Kit
AFP(ng/mL) / Intra-assay Inter-assay
Mean S.D CV% Mean S.D CV%
20 / 21 1.2 5.7 / 22 1.3 5.9
50 / 53 3.5 6.6 / 52 3.8 7.3
200 / 202 6.2 6.2 / 205 11.1 5.4

4. Linearity: The high concentration was diluted with the low concentration to the following final percentages; 100%, 75%, 50%, 25%, 10%, 5% and 0%. Sample was assayed in triplicate in one analytical run at each AFP level. The coefficient of linear regression was R=0.997. Linearity of i-CHROMATM AFP TestKit was 5~350ng/mL.

5. Comparability:The AFP concentrations of 30clinical specimens were quantified independently with i-CHROMATM AFP TestKit and Axsym (Abbott) automatic analyzer. While the whole blood was used for i-CHROMATM AFP TestKit, the serums were used for Abott Axsym. The test results were comparedand their compatibilities were investigated with linear regression and correlation of coefficient (R). i-CHROMATM AFP Testwas comparable well to other method (R=0.989).

REFERENCES

1. Tartarinov, Y.S. Detection of embryospecific alpha-globulin in the blood sera of patients with primary liver tumor. Vopr. Med. Khim. 10:90-91 (1964).
2. Mcintire, K.R., Waidmann, T.A., Moertel, C.G. and Go, V.L.W. Serum alpha-fetoprotein in patients with neoplasms of the gastrointestinal tract. Cancer Res. 35:991-996 (1975).
3. Javadpouf, N., Mcintire, K.R. and Waidmann, T.A. Human chorionic gonadotropin (HCG) and alpha-fetoprotein (AFP) in sera and tumor cells of patients with testicular seminoma. Cancer 42:2768-2772. (1978).
4. Chen, D.S. and Sung, J.L. Relationship of Hepatitis B Surface Antigen to serum alpha-fetoprotein in nonmalignant diseases of the lever. Cancer 44:984-992 (1979).
5. Rhoslati,E. and Seppala, M. studies of carcinofetal proteins: Physical and Chemical Properties of Human alpha-fetoprotein. Int. J. Cancer 7:218-225 (1971).
6. Abelev,G.I. Alpha-fetoprotein in oncogenesis and its association with malignant tumors. Adv. Cancer Res. 7:295-358 (1971).
7. Wespic, H.C. Alpha-fetoprotein: its quantification and relationship to neoplastic disease, ppp 115-129 In Alpha-fetoprotein, Laboratory Procedures and Clinical Applications, Kirkpatirck, A. and Nakamuram R (eds.), Masson Publishing, New York (1981) After Radical Prostatectomy. J. Urol. 142:1082-90 (1989).

Rev. 05_100513_M