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|Year : 1993 | Volume
| Issue : 3 | Page : 130-1
Comparative evaluation of coagglutination and latex agglutination test (Rotalex kit) for detection of rota virus.
MS Mathur, GG Bhave
Dept of Microbiology, Seth GS Medical College, Parel, Bombay, Maharashtra.
M S Mathur
Dept of Microbiology, Seth GS Medical College, Parel, Bombay, Maharashtra.
Source of Support: None, Conflict of Interest: None
Coagglutination test was compared with commercially available latex agglutination test (Rotalex kit) for detection of rota virus in faecal samples from clinically suspected cases of viral gastroenteritis. Out of 80 test samples 16 (20%) and 20 (25.3%) were positive for rota virus antigen by Rotalex kit and coagglutination test respectively. All the 40 controls were negative for viral antigen by Rotalex kit and only one gave positive result by coagglutination test. Coagglutination test was found to be economical, sensitive and specific for screening and rapid diagnosis of Rota virus diarrhoea.
Keywords: Agglutination Tests, Child, Child, Preschool, Diarrhea, microbiology,Feces, microbiology,Female, Gastroenteritis, microbiology,Human, Infant, Latex Fixation Tests, Male, Reagent Kits, Diagnostic, Rotavirus, isolation &purification,Rotavirus Infections, microbiology,
|How to cite this article:|
Mathur M S, Bhave G G. Comparative evaluation of coagglutination and latex agglutination test (Rotalex kit) for detection of rota virus. J Postgrad Med 1993;39:130
|How to cite this URL:|
Mathur M S, Bhave G G. Comparative evaluation of coagglutination and latex agglutination test (Rotalex kit) for detection of rota virus. J Postgrad Med [serial online] 1993 [cited 2020 Jul 11];39:130. Available from: http://www.jpgmonline.com/text.asp?1993/39/3/130/617
Rota virus has been reported as the major cause of acute gastroenteritis in young children,,,. During infection rota virus particles are excreted in large number in the faeces, and can be easily observed and identified by electron microscopy of stool samples. Though this is the most specific method for detecting rota virus infection, facility of electron microscope is not available in most of the laboratories, hence one has to use other methods like counter immunoelectroosmophoresis (Cl EP), RIA (radio-immuno assay), ELISA (enzyme linked immunosorbent assay), passive haemagglutination (PHA), latex agglutination test (LAT), and coagglutination for detection of rota virus in stool samples.
In the present study an attempt has been made to standarditse coagglutination technique for detection of rota virus in stool samples and compare the results with the commercially available Rotalex kit from Orion Diagnostica, Helsinki, Finland.
A total of 80 faecal samples from cases of acute gastroenteritis from children of age group 1-8 years and 40 non-diarrhoeal controls from paediatric age group were included in the present study. Faecal samples were collected before starting anti- diarrhoeal therapy and preserved at -20' C until use.
1. Rotalex test: Faecal samples were processed as per instructions given in the manual provided with Rotalex kit. In short, stool samples were diluted 1:10 with Rotalex buffer and mixed with vortex mixer and the suspension is allowed to stand for about 30 minutes at room temperature followed by centrifugation for 30 minutes at 3000 rpm. Then 2 separate drops of 50 ?l each from the above supernatent were put on a slide. Rotalex reagent was added to one drop and to the other drop Rotalex control latex reagent was added. The slide was tilted back and forth for two minutes. Development of agglutination in Rotalex reagent was treated as positive. If agglutination was observed in the drop containing Rotalex control latex reagent, the sample was not included in the study. Positive and negative controls were run with each test.
2. Coagglutination test: Protein A reagent was prepared by coating Staphylococcus aureus, Cowan 1 (NCTC 8530) strain with bovine antisera against rota virus antigen (Wellcome Diganostics, UX). The procedure adopted was as follows: Cowan 1 protein A rich stain of Staphylococcus was grown on Muller Hinton agar. Cells were harvested in phosphate buffered saline (PBS) pH 7.4 and washed thrice in the same buffer and the suspension was heated at 80?C for 1 hour. Then it was centrifuged at 2000 rpm for 30 minutes, supernatent was discarded and the pellet was resuspended in PBS pH 7.4, 10% w/V. The cells were again washed thrice and stored in 10% suspension in PBS at 4?C. One ml of 10% Staphylococcal suspension was mixed with 0.1 ml of neat rota virus antiserum (Wellcome Diagnostics, U.K.) and incubated for 1 hour at 37?C in water bath. This reagent is stable for 4 weeks at 4?C.
Stool samples were processed in the same way in Rotalex test. Fifty microlitre of sensitised protein reagent was added to the 50 ?l of supernatent from faecal suspension taken on a slide and coagglutination was observed with the naked eye within 2 minutes. A known positive and negative control was accompanied with each batch of tests.
Of 80 test samples, 16 (20%) were positive for rota virus antigen by Rotalex kit and 20 (25.3%) by coagglutination test. All the 40 controls were negative for rota virus antigen by Rotalex kit and only one gave positive result by coagglutination test. Thus 4 test samples and 1 control sample gave positive result with coagglutination test but were negative for antigen as detected by Rotalex test.
Rota virus is the major aetiological agent causing 40-60% of childhood diarrhoea in different parts of the world (WHO). In India, rota virus incidences have been reported to be 17-32% from studies in Chandigarh, Vellore and Delhi. Among different techniques used for detection of rota virus antigen in faecal samples RIA, ELISA, electron microscopy are very sensitive and specific but require sophisticated equipment,. So the need has always been felt to develop cheaper tests like latex agglutination and coagglutination tests. In the present study coagglutination test gave positive results in 25% of the test samples and Rotalex test in 20% of the cases, proving that coagglutination is more sensitive than Rotalex test. In control group coagglutination test was positive only in 2.5% of cases showing that coagglutination test correlates well with Rotalex test. The Rotalex kits are to be imported and are costly. Therefore coagglutination test is a good substitute for mass screening and quick diagnosis of rota virus diarrhoea.
We are thankful to Dr (Mrs) PM Pai, Dean, Seth GS Medical College and King Edward Memorial Hospital, Mumbai 400 012 for her kind permission to publish this paper.
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