BODITECH MED INC. I-CHROMA MICROALBUMIN-25

BODITECH MED INC. i-CHROMA MICROALBUMIN-25

REFERENCES

1. Rowe DJF, Dawnay A, Watts GF. Microalbuminuria indiabetes mellitus: review and recommendations for the measurement of albumin in urine. Ann Clin Biochem 1990; 27: 297-312.

2. Doumas BT, PetersT. Serum and urine albumin: a progress report on their measurement and clinical significance. Clin Chim Acta 1997; 258:3-20.

3. Mogensen CE. Microalbuminuria, a marker for organ damage. 1993. London: Science Press.

4. Waugh J, Kilby M, Lambert P, Bell SC, Blackwell CN, Shennan A, et al. Validation of the DCA 2000 microalbumin:creatinine ratio urinanalyzer for its use in pregnancy and preeclampsia.Hypertens Pregnancy 2003; 22(1): 77-92.

5. Mogensen CE, Christnesen CK. Predicting diabetic nephropathy in insulin dependent diabetes. New Eng J Med 1984; 311:89-93.

6. Viberti GC, Hill RD, Jarrett RJ. Microalbuminuria as a predictor of clinical nephropathy in insulin dependent diabetes mellitus. Lancet 1982;I: 1430-2.

7. Mathiesesen ER, Ronn B, Jensen T, and Deckert, T. Relationship between blood pressure and urinary excretion in the development of microalbuminurea. Diabetes 1990; 39:245-9.

8. Brooks DE, Devine DV, Harris PC, et al. RAMP(TM): A rapid, quantitative whole blood immunochromatographic platform for point-of-care testing.Clin Chem 1999;45:1676-1678.

9. Oh SW, Moon JD, Park SY, et al. Evaluation of fluorescence hs-CRP immunoassay for point-of –care testing. Clin Chim Acta 2005; 356:172-177.

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Revision No: 08

Date of last revision: May13, 2010.

i-CHROMATM Microalbumin

ImmunoAssay for Quantitative Measurement of Microalbumin in Human urine with i-CHROMATM Reader System

INTENDED USE

The i-CHROMATM Microalbuminalong with i-CHROMATM Reader is a fluorescence immunoassay that measures albumin in urine. The test is used as an aid to predict diabetic nephropathy as well as future cardiovascular diseases (CVD).

INTRODUCTION

A MAU(microalbumin) test evaluates urine for the presence of a protein called albumin1. Albumin is normally found in the blood and filtered by the kidneys2. When the kidneys are working properly, albumin is not present in the urine. However, when the kidneys are damaged, small amounts of albumin leak into the urine. This condition is called microalbuminuria1, 2,3,4.

Microalbuminuria is most frequently caused by kidney damage from diabetes. However, many other conditions can lead to kidney damage, such as high blood pressure, heart failure, cirrhosis, or systemic lupus erythmatosus(SLE). If early kidney damage is not treated, larger amounts of albumin and protein may leak into the urine5,6. This condition is called macroalbuminuria or proteinuria. When the kidneys spill protein, it can mean serious kidney damage is present. This can lead to chronic kidney disease. A microalbumin urine test can be done on a sample of urine collected randomly (usually after the first time you urinate in the morning), a sample collected over a 24-hour period, or a sample collected over a specific period of time, such as 4 hours or overnight7.

PRINCIPLE

The i-CHROMATM Microalbuminis based on fluorescence immunoassay technology8, 9. The i-CHROMATM Microalbumin uses a sandwich immunodetection method, such that by mixing detector buffer with urine specimen in test vial, the fluorescence-labeled detector anti-albumin antibodyin buffer binds to albumin antigen in urinespecimen. 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 albumin are captured to anti-albumin sandwich pair antibody that has been immobilized on test strip. Thus the more albuminantigen is inurine specimen, the more complexes are accumulated on test strip. Signal intensity of fluorescence of detector antibody reflects amount of albumin captured and is microprocessed from i-CHROMATM Reader to show albumin concentration in urine specimen. The default result unit of i-CHROMATM MAU is displayed as an mg/L from i-CHROMATM Reader. The working range of i-CHROMATM MAU system is 2 - 300 mg/L.

* Reference Value : 18mg/L

COMPOSITION OF REAGENTS

Thei-CHROMATM Microalbumin Kit consists of Test Device, ID Chip, and Detector Buffer. Test Device is individually sealed with a desiccant in aluminum pouch, and Detector Buffer is dispensed individually in a tube and a bottle containing the predispensed tubes is delivered separately from Test Device in a styrofoam box filled with ice pack.

•Test Device contains a test strip in which mouse anti-albumin monoclonal antibody andrabbit IgG have been immobilized on the test and on the control line of strip, respectively.

• Detector Buffer, predispensed in a tube, contains fluorescence-labeled anti-human albumin (mouse monoclonal, 3 µg/mL), fluorescence-labeled anti-rabbit IgG (500 ng/mL), 1% gelatin as a stabilizer, and 0.1% 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 MAU -25

• Don’t use Test Device if its lot number does not match with EEPROM card number that is inserted onto the instrument.

• Thei-CHROMATM Microalbumin is only operational in thei-CHROMATM Reader. Andtests should be applied by trainedstaff working in the laboratories where 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 Microalbumin Device 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.

•Thei-CHROMATM Microalbumin Deviceand Reader should be used away from vibration and magnetic field. During normal usage, i-CHROMATM Microalbumin 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.

• Urine 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..

•Thei-CHROMATM Microalbumin should not be used asabsolute evidence fordiabetic nephropathy and high risk of CVD. The results should be interpreted by the physician along with clinical findingsand other laboratory test results.

•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

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

• Storei-CHROMATM Microalbumin Device at 4°-30°C in its sealed pouch. The i-CHROMATM Microalbumin Deviceis 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 human urine samples.

•The urine 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. .

MATERIALS PROVIDED

BodiTech Med Incorporated i-CHROMATM Microalbumin

Catalog REF Catalog No. i-CHROMATM MAU -25

Kit contains:

Test Devices 25T/box

ID Chip 1/box

Detection Buffer 25 tubes/pouch

MATERIALSREQUIRED BUT NOT PROVIDED

i-CHROMATM Reader REF Catalog No.FR-203

Thermal Printer

Transfer pipette (10, 75µL size)

PROCEDURE

• Image of the test kit

window sample well

1. Check/insert ID chip onto instrument. Make sure that the Test Device lot # matches with ID chip #.
2. Set a Test Device on a dust-free clean place.
3. Take out a tube ofDetectionBufferfrom the bottles in a refrigerator and leave them at room temperature.
4. Draw 10µL of urinewith a transfer pipette and add it to the tube containing DetectionBuffer.
5. Mix well the specimenwith DetectionBuffer by inverting repeatedly the tube.
6. Take 75µL of sample mixture and load it onto the well of a disposable Test Device.
7. Leave the Test Device at room temperature for 12 min before inserting the device into the holder.
8. To start scanning, insert test device onto the holder of i-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 ofi-CHROMATM Reader or tear out the print for record.

Refer to i-CHROMATMReader Operation Manual for the complete instructions on use of the Test. REF Catalog No. FR- 203

RESULT

Thei-CHROMATM Reader calculates MAUtest results automatically and displays concentration of microalbumin in urine on the LCD as form of mg/L.For further information, refer to the Operation Manual for the i-CHROMATM Reader.

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 usingWako albumin 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 Incorporated’s Technical Services for assistance.

Procedure Control

Each i-CHROMATM Microalbumin Device 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 Readerindicating that the test should berepeated.

LIMITATIONS OF THE PROCEDURE

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

• The false positive results include cross-reactions withsome components of urine from individual to antibodies; and non-specific adhesionof some components in human urine 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 albumin 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.

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

PERFORMANCE CHARACTERISTICS

1. Analytical Sensitivity: Analytical sensitivity means the lowest concentration of Microalbumin that the test system can detect with CV<10%. Analytical sensitivity of i-CHROMATMMAU Test was determined by testing 10 times with three lots of reagents. Analytical sensitivity of i-CHROMATMMAU Test system was 2 mg/L.

2. Specificity:Other bio-molecules, such as Hb, CEA, PSA, AFP, ALP, CRP, Troponin I, Myoglobin component were added to test specimen with much higher level than their physiological level in normal blood. There was no significant interference with the Microalbuminmeasurement, nor was their any significant assay cross-reactivity with those bio-molecules tested.

3. Imprecision: For the intra-assay imprecision, 10 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 Microalbumin concentration.

Imprecision of i-CHROMATM MAUTest Kit
Microalbumin(mg/L) / Intra-assay Inter-assay
Mean S.D CV% Mean S.D CV%
25 / 24.3 1.0 4.32 / 24.5 1.5 6.6
100 / 100.2 5.5 5.48 / 100.7 7.6 7.6
200 / 200.6 6.5 3.25 / 200.0 13.8 6.9

4. Linearity: The high pool (100 mg/L) was diluted with the very low pool (~0.10 mg/L) to the following final percentages; 100%, 50%, 25%, 12.5%, 6.2%, 3.1% and 1.5%. Sample was assayed in triplicate in one analytical run at each Microalbumin level. The coefficient of linear regression was R2=0.99.

5. Comparability:The Microalbumin concentrations of 100clinical specimens were quantified independently with i-CHROMATMMAUand DCA2000 according to their standard test procedure. The test results were comparedand their compatibilities were investigated with linear regression and correlation of coefficient (R2). i-CHROMATMMAUwas comparable well to other method (R2=0.99).

Rev. 08_100513_M