CLIA COMPLEXITY:  Waived

I. INTENDED USE

The Clearview Mono test is a rapid chromatographic immunoassay for the qualitative detection of Infectious Mononucleosis heterophile antibodies in whole blood to aid in the diagnosis of Infectious Mononucleosis.   For professional in vitro diagnostic use only. 

II. PRINCIPLE

The Clearview Mono test is a qualitative membrane strip based immunoassay for the detection of IM heterophile antibodies in whole blood. In this test procedure, bovine erythrocyte extracted antigen is coated on the test line region of the strip. The sample reacts with bovine erythrocyte extracted antigen coated particles that have been applied to the label pad. This mixture migrates chromatographically along the length of the test strip and interacts with the coated bovine erythrocyte extracted antigen. If the sample contains IM antibodies, a colored line will appear in the test line region indicating a positive result. If the sample does not contain IM heterophile antibodies, a colored line will not appear in this region indicating a negative result. To serve as a procedural control, a colored line will always appear at the control line region, indicating that proper volume of specimen has been added and membrane wicking has occurred.

III. SPECIMEN

IV. REAGENT AND EQUIPMENT

A.     Reagents and Materials Provided

1. 30 Test devices
2. Disposable sample tubes
3. Disposable heparinized capillary tubes and dispensing bulb
4. ositive control (Diluted human plasma containing IM heterophile antibodies, 0.09% sodium azide)
5. Negative control (Diluted human plasma, 0.09% sodium azide)
6. Sample buffer
7. Package insert              

B.     Materials Required but not Provided

1. Sample collection container (for venipuncture whole blood)
2. Lancet (for fingerstick whole blood only)
3. Timer

V. STORAGE AND STABILITY

The kit can be stored at room temperature or refrigerated (2°-30°C). The test devices must remain in the sealed pouch until use. The test device is stable through the expiration date printed on the sealed pouch. The test device must remain in the sealed pouch until use. DO NOT FREEZE. Do not use beyond the expiration date.

VI. QUALITY CONTROL:

Internal Procedural Controls:

• Internal procedural controls are included in the test.
  • A red line appearing in the control region (C) is an internal positive procedural control. It confirms sufficient sample volume and correct procedural technique.
  • A clear background is an internal negative background control. If the test is working properly, the background in the result area should be white to light pink and not interfere with the ability to read the test result.

External Procedural Controls:

• Using the positive or negative external controls in place of a patient sample, add 1 drop of positive or negative control solution to the sample well (S) of a new test device, then add 1 drop of Sample Buffer. Start the timer. Continue with Step 3 in the Directions For Use Section. Positive and negative external controls be run with each new lot.

Remedial Actions

• When correct control results are not obtained, do not report patient results.  Contact Point-of Care-Testing Coordinator phone: 444-2320 or
• Inverness Medical Technical Services at 800-637-3717.

VII.  IMPLEMENTATION Test Procedure

A. Specimen Collection and Handling:

1. To collect Venipuncture Whole Blood samples:  Collect anti-coagulated blood sample (sodium or potassium heparin, sodium or potassium EDTA, sodium or potassium citrate and sodium oxalate) following standard laboratory procedures.
2. To collect Fingerstick Whole Blood samples:
   1. Wash the patient’s hand with soap and warm water or clean with an alcohol swab. Allow to dry.
   2. Massage the hand without touching the puncture site by rubbing down the hand towards the fingertip of the middle or ring finger.
   3. Puncture the skin with a sterile lancet. Wipe away the first sign of blood.
   4. Gently rub the hand from wrist to palm to finger to form a rounded drop of blood over the puncture site.
   5. Touch the end of the capillary tube to the blood until filled to the line; avoid air bubbles.
   6. Place the bulb onto the top end of the capillary tube.
   7. Squeeze the bulb to dispense the whole blood

B. Test Procedure:

1. Allow the test device, sample, buffer and controls to reach room temperature (15° - 30°C) before testing.
2. Remove the test device from the foil pouch and use it as soon as possible. For best results, perform the test immediately after opening the foil pouch.
3. Place the device on a clean level surface.
4. For Whole Blood (Venipuncture) samples:

• For Whole Blood (Fingerstick) samples:

Fill dropper with sample. Hold the dropper upright, and add 2 drops of whole blood (about 50 µL) to the sample well of the test device. Add 1 drop of Sample Buffer to the sample well. Start the timer. Wait for the red line(s) to appear. The result should be read at 5 minutes. The background should be clear before the result is read. Record the test result, and procedural control result on the Log sheet Record the test result electronically in Meditech. View procedure

• For Whole Blood (Fingerstick) samples:

Add one capillary tube of blood (about 50 µL) to the sample well(s) of the test device. Add 1 drop of Sample Buffer to the sample well. Start the timer. Wait for the red line(s) to appear. The result should be read at 5 minutes. The background should be clear before the result is read. Record the test result, and procedural control result on the Log sheet Record the test result electronically in Meditech.

VIII. DOCUMENTATION:

Record the test result, and procedural control result on the Log sheet
Record the test result electronically in Meditech.

IX. NOTES AND PRECAUTIONS:

1. Low titers of IM heterophile antibodies might result in a weak line appearing in the test line region (T) after a long period of time.  Do not read the result after 10 minutes.
2. Testing should ideally be performed immediately after the samples have been collected. Do not leave the samples at room temperature for prolonged periods. Whole blood collected by venipuncture should be stored at 2°- 8°C if the test is to be run within 2 days of collection.  Whole blood collected by fingerstick should be tested immediately.  Do not freeze whole blood samples.
3. For professional in vitro diagnostic use only. Do not use after expiration date.
4. Do not eat, drink or smoke in the area where the specimen samples and kits are handled.
5. The positive and negative controls contain human plasma. Handle controls and all specimen samples as if they contain infectious agents. Observe established precautions against microbiological hazards throughout all procedures and follow the standard procedures for proper disposal of specimen samples.
6. The positive and negative controls contain sodium azide as a preservative, which may form potentially explosive metal azide if it reacts with lead or copper plumbing. Large quantities of water should be used to flush discarded controls down a sink.
7. Wear protective clothing such as laboratory coats, disposable gloves and eye protection when specimen samples are assayed.
8. Humidity and temperature can adversely affect results.
9. The dispensing bulb used with the capillary tubes to add fingerstick whole blood to the device may contain trace amounts of latex, which may cause an allergic reaction in some individuals.
10. Grossly hemolysed samples will yield invalid results. Strictly follow the Package Insert instructions to obtain accurate results.

X.    INTERPRETATION:

POSITIVE*: Two distinct red lines appear. One line should be in the control line region (C) and another line should be in the test line region (T). A positive result means that IM heterophile antibodies were detected in the sample.

*NOTE: The shade of the red color in the test line region (T) will vary based on the amount of IM heterophile antibodies in the sample. Any shade of red in the test line region (T) should be considered positive.

NEGATIVE: One red line appears in the control line region (C). No apparent red or pink line appears in the test line region (T). A negative result means that IM heterophile antibodies were not found in the sample or are below the detection limit of the test.

INVALID: No line appears in the control line region (C). Review the procedure and repeat the test with a new test device. If the problem persists, discontinue using the test kit immediately and contact Inverness Medical Technical Support at (800) 637-3717.

XI.      LIMITATIONS:

1. The Clearview Mono test (Whole Blood) is for in vitro diagnostic use only. The test should be used for the detection of IM heterophile antibodies in whole blood samples only. Neither the quantitative value nor the rate of increase in Infectious Mononucleosis antibody concentration can be determined by this qualitative test.
2. The Clearview Mono test (Whole Blood) will only indicate the presence of IM heterophile antibodies in the sample and should not be used as the sole criteria for the diagnosis of Mononucleosis infection.
3. Grossly hemolysed samples will yield invalid results. Strictly follow the Package Insert instructions to obtain accurate results.
4. As with all diagnostic tests, all results must be interpreted together with other clinical information available to the physician.
5. Patients under 18 years of age were not included in the clinical study.  A heterophile antibody response is observed in approximately 80-90% of adults and children with EBV-caused IM, and observed in approximately 50% of children under four years of age with EBV-caused IM.

XII.        EXPECTED VALUES:

1. Epstein-Barr virus infection during adolescence or young adulthood causes Infectious Mononucleosis 35% to 50% of the time.^{1, 5} 
2. The incidence of EBV-associated Infectious Mononucleosis in the USA has been estimated at 45 per 100,000 and is highest in adolescent and young adults about 2 out of 1,000. No seasonal pattern of EBV infection exists. The incubation period is 10 to 60 days, though 7 to 14 days is common for children and adolescents.

XIII.        INTERFERENCE:

No interference with the Clearview Mono test results was observed in samples containing high levels of hemoglobin (up to 1,000 µg/dL), bilirubin (up to 1,000 mg/dL) and human serum albumin (up to 2,000 mg/dL).  The test results were also unaffected when the hematocrit was altered ranging from 20% to 60% and when icteric and lipemic samples were tested.

XIV.       REFERENCES:

1. Pediatr Clin North Am 1997 Dec;44(6):1541-56
2. Omori, M. 2002 Mononucleosis. http://www.emedicine.com/EMERG/topic319.htm
3. Linde A. 1996. Scand J Infect Dis Suppl. 100:83-8
4. Papesch, M. & Watkins, R. 2001 Clin. Otolaryngol. 26, 3-8
5. CDC National Center for infectious Diseases: EBV & IM: www.cdc.gov/ncidod/diseases/ebv.htm
6. Lennette, ET. Epstein-Barr Virus. 1991, Manual of Clinical Microbiology, 5th Edition. American Society for Microbiology, Washington DC. 847-852
7. Cearview Mono Whole Blood package insert :03/07 #1155897202, product# 92401