ACCU-TELL ROTAVIRUS/ADENOVIRUS COMBO RAPID TEST

CATALOG

Catalog No.                     Product Name

ABT-IDT-B64                   Adeno/Rota Combo Cassette
INTENDED USE
Accu-Tell^® Rotavirus/Adenovirus Combo Rapid Test Device (Feces) is a rapid visual immunoassay for the qualitative, presumptive detection of rotavirus and adenovirus in human fecal specimens. This kit is intended for use as an aid in the diagnosis of rotavirus and adenovirus infection.

INTRODUCTION
Rotavirus is the most common agent responsible for acute gastroenteritis, mainly in young children. Its discovery in 1973 and its association with infantile gastroenteritis represented a very important advancement in the study of gastroenteritis not caused by acute bacterial infection. Rotavirus is transmitted by oral-fecal route with an incubation period of 1-3 days. Although specimens collected within the second and fifth day of the illness are ideal for antigen detection, rotavirus may still be found while diarrhea continues. Rotaviral gastroenteritis may result in mortality for populations at risk such as infants, the elderly and immunocompromised patients. In temperate climates, rotavirus infections occur mainly in the winter months. Endemics as well as epidemics affecting some thousand people have been reported. With hospitalized children suffering from acute enteric disease, up to 50% of the analyzed specimens were positive for rotavirus. The viruses replicate in the cell nucleus and tend to be host species-specific producing a characteristic cytopathic effect (CPE). Because rotavirus is extremely difficult to culture, it is unusual to use isolation of the virus in the diagnosis of infections. Instead, a variety of techniques have been developed to detect rotavirus in feces. Acute diarrheal disease in young children is a major cause of morbidity worldwide and is a leading cause of mortality in developing countries. Research has shown that enteric adenoviruses, primarily Ad40 and Ad41, are a leading cause of diarrhea in many of these children, second only to the rotaviruses. These viral pathogens have been isolated throughout the world, and can cause diarrhea in children year round. Infections are most frequently seen in children less than two years of age, but have been found in patients of all ages. Further studies indicate that adenoviruses are associated with 4-15% of all hospitalized cases of viral gastroenteritis.
Rapid and accurate diagnosis of adenovirus-related gastroenteritis is helpful in establishing the etiology of gastroenteritis and related patient management. Other diagnostic techniques such as electron microscopy (EM) and nucleic acid hybridization are expensive and labor-intensive. Given the self-limiting nature of adenovirus infection, such expensive and labor-intensive tests may not be necessary.

PRINCIPLE
Accu-Tell^® Rotavirus/Adenovirus Combo Rapid Test Device (Feces) detects rotavirus and adenovirus through visual interpretation of color development on the internal strip. Anti-rotavirus antibodies and anti-adenovirus are immobilized on the test region of the membrane. During testing, the specimen is allowed to react with anti-rotavirus antibodies and anti-adenovirus antibodies conjugated to colored particles and precoated onto the sample pad of the test. The mixture then migrates through the membrane by capillary action and interacts with reagents on the membrane. If there is sufficient rotavirus in the specimen, a colored band will form at the R region of the membrane. Likewise, if there is sufficient adenovirus in the specimen, a colored band will form at the A region of the membrane. The presence of colored band(s) indicates a positive result, while their absence indicates a negative result. The appearance of a colored band at the control region serves as a procedural control, indicating that the proper volume of specimen has been added and membrane wicking has occurred.

KIT COMPONENTS
1.  Individually packed test devices
2.  Each test contains colored conjugates and reactive reagents precoated at the corresponding regions
3.  Specimen collection cards For specimens collection
4.  Specimen dilution tube with buffer 0.1 M Phosphate buffered saline (PBS) and 0.02% sodium azide
5.  Disposable pipettes For collection of liquid specimens
6.  Package insert For operating instructions

PRECAUTIONS
1.  For professional in vitro diagnostic use only.
2.  Do not use after the expiration date indicated on the package. Do not use the test if the foil pouch is damaged. Do not reuse tests.
3.  This kit contains products of animal origin. Certified knowledge of the origin and/or sanitary state of the animals does not completely guarantee the absence of transmissible pathogenic agents. It is therefore, recommended that these products be treated as potentially infectious, and handled by observing usual safety precautions (e.g., do not ingest or inhale).
4.  Avoid cross-contamination of specimens by using a new specimen collection container for each specimen obtained.
5.  Read the entire procedure carefully prior to testing.
6.  Do not eat, drink or smoke in any area where specimens and kits are handled. Handle all specimens as if they contain infectious agents. Observe established precautions against microbiological hazards throughout the procedure and follow standard procedures for the proper disposal of specimens. Wear protective clothing such as laboratory coats, disposable gloves and eye protection when specimens are assayed.
7.  The specimen dilution buffer contains sodium azide, which may react with lead or copper plumbing to form potentially explosive metal azides. When disposing of specimen dilution buffer or extracted samples, always flush with copious quantities of water to prevent azide buildup.
8.  Do not interchange or mix reagents from different lots.
9.  Humidity and temperature can adversely affect results.
10.  Used testing materials should be discarded according to local regulations.

STORAGE AND STABILITY
1.  The kit should be stored at 2-30°C until the expiry date printed on the sealed pouch. • The test must remain in the sealed pouch until use.
2.  Do not freeze.
3.  Care should be taken to protect the components of this kit from contamination. Do not use if there is evidence of microbial contamination or precipitation. Biological contamination of dispensing equipment, containers or reagents can lead to false results.

SPECIMEN COLLECTION AND STORAGE
1.  Accu-Tell^® Rotavirus and Adenovirus Combo Rapid Test Device (Feces) is intended for use with human fecal specimens only.
2.  Viral detection is improved by collecting the specimens at the onset of symptoms. It has been reported that the maximum excretion of rotavirus in the feces of patients with gastroenteritis occurs 3-5 days after onset of symptoms. If the specimens are collected long after the onset of diarrheic symptoms, the quantity of antigen may not be sufficient to obtain a positive result, or the antigens detected may not be linked to the diarrheic episode.
3.  Perform testing immediately after specimen collection. Do not leave specimens at room temperature for prolonged periods. Specimens may be stored at 2-8°C for up to 72 hours.

PROCEDURE
Bring specimens to room temperature prior to testing. If specimens are to be shipped, pack them in compliance with all applicable regulations for transportation of etiological agents.
Bring tests, specimens, buffer and/or controls to room temperature (15-30°C) before use.
1. Specimen collection and pre-treatment:
1)  Use the specimen collection cards provided in the kit for specimen collection. Follow the instructions on the card. Other clean, dry containers may also be used for specimen collection. Best results will be obtained if the assay is performed within 6 hours after collection.
2)  For solid specimens: Unscrew and remove the dilution tube applicator. Be careful not to spill or spatter solution from the tube. Collect specimens by inserting the applicator stick into at least 3 different sites of the feces to collect approximately 50 mg of feces (equivalent to 1/4 of a pea).
For liquid specimens: Hold the pipette vertically, aspirate fecal specimens, and then transfer 2 drops (approximately 50 μL) into the specimen collection tube containing the extraction buffer.
3)  Replace the applicator back into the tube and screw the cap tightly. Be careful not to break the tip of the dilution tube.
4)  Shake the specimen collection tube vigorously to mix the specimen and the extraction buffer. Specimens prepared in the specimen collection tube may be stored for 6 months at -20°C if not tested within 1 hour after preparation.
2. Testing
1)  Remove the test from its sealed pouch, and place it on a clean, level surface. Label the test with patient or control identification. For best results, the assay should be performed within one hour.
2)  Using a piece of tissue paper, break the tip of the dilution tube. Hold the tube vertically and dispense 2 drops of solution into the specimen well (S) of the test device.
Avoid trapping air bubbles in the specimen well (S), and do not add any solution to the result window.
As the test begins to work, color will migrate across the membrane.
3. Wait for the colored band(s) to appear. The result should be read at 10 minutes. DO NOT INTERPRET THE RESULT AFTER 20 MINUTES.
Note: If the specimen does not migrate due to the presence of particles, centrifuge the extracted specimen contained in the extraction buffer vial.
Collect 80 μL of supernatant, dispense into the specimen well (S) of a new test device and start again, following the instructions mentioned above.

INTERPRETATION OF RESULTS
POSITIVE:
Rotavirus Positive: A colored band appears in the control band region (C) and another colored band appears in the R band region.
Adenovirus Positive: A colored band appears in the control band region (C) and another colored band appears in the A band region.
Rotavirus and Adenovirus Positive: A colored band appears in the control band region (C) and two other colored bands appear in the A band region and R band regions, respectively.
NEGATIVE:
Only one colored band appears, in the control band region (C). No band appears in either test band region (A/R).
INVALID:
Control band fails to appear. Results from any test which has not produced a control band at the specified reading time must be discarded. Please review the procedure and repeat with a new test. If the problem persists, discontinue using the kit immediately and contact your local distributor

NOTE:
1.  The intensity of color in the test regions (A/R) may vary depending on the concentration of analytes present in the specimen. Therefore, any shade of color in the test regions should be considered positive. Note that this is a qualitative test, and cannot determine the concentrations of analytes in specimens.
2.  Insufficient specimen volume, incorrect operating procedure or expired tests are the most likely reasons for control band failure.

QUALITY CONTROL
1.  Internal procedural controls are included in the test. A colored band appearing in the control region (C) is considered an internal positive procedural control, confirming sufficient specimen volume and correct procedural technique.
2.  External controls are not supplied with this kit. It is recommended that positive and negative controls be tested as a good laboratory practice to confirm the test procedure and to verify proper test performance.

LIMITATIONS OF THE TEST
1. The Rotavirus/Adenovirus Rapid Test Device (Feces) is for professional in vitro diagnostic use, and should only be used for the qualitative detection of rotavirus.
2. As with all diagnostic tests, a definitive clinical diagnosis should not be based on the results of a single test, but should only be made by the physician after all clinical and laboratory findings have been evaluated.
3. If the test result is negative and clinical symptoms persist, additional testing using other clinical methods is recommended. A negative result does not at any time rule out the possibility of rotavirus or adenovirus infection, as virus particles may be present below the minimum detection level of the test.

PERFORMANCE CHARATERISTICS

Specificity:
Cross reactivity with the following organisms has been studied at 1.0 x 109 organisms/mL. The following organisms produced negative results with Accu-Tell® Rotavirus/Adenovirus Combo Rapid Test Device (Feces).
Staphylococcus aureus
Proteus mirabilis
Neisseria gonorrhea
Pseudomonas aeruginosa
Acinetobacter spp
Group B Streptococcus
Enterococcus faecalis
Salmonella choleraesius
Proteus vulgaris
Group C Streptococcus
Gardnerella vaginalis
Enterococcus faecium
Klebsiella pneumoniae
Acinetobacter calcoaceticus
Hemophilus influenzae
Branhamella catarrhalis
Escherichia coli
Neisseria meningitides
Candida albicans
Chlamydia trachomatis

LITERATURE REFERENCES
1. Wadell G. Adenoviridae: The adenoviruses. In: Lennette EH, Halonen P, Murphy FA, eds. Laboratory diagnosis of infectious diseases: Principles and practices, Vol II. Viral, rickettsial, and chlamydial diseases. New York, NY: Springer-Verlag;
1988: 284-300.
2. Wilhelmi I, Roman E, Sánchez-Fauquier A. Viruses causing gastroenteritis. Clin Microbiol Infect. 2003 Apr; 9(4): 247-62.
3. Cubitt WD. Rotavirus infection: an unexpected hazard in units caring for the elderly. Geriat Med Today. 1982; 1:33-38.
4. Wood DJ, Bijlsma K, de Jong JC, Tonkin C. Evaluation of a commercial monoclonal antibody-based enzyme immunoassay for detection of adenovirus types 40 and 41 in stool specimens. J Clin Microbiol. 1989 Jun; 27(6): 1155-8.
5. Thomas EE, Roscoe DL, Book L, Bone B, Browne L, Mah V. The utility of latex agglutination assays in the diagnosis of pediatric viral gastroenteritis. Am J Clin Pathol. 1994 Jun.

The above information is just for reference. The actual technical specifications are subject to the insert provided with the product.