ABSTRACT
BACKGROUND & OBJECTIVES: Rotavirus is the major cause of gastroenteritis in infants and young children all over the world. The objective of the study was to develop a rapid ELISA for the diagnosis of rotavirus infection in children hospitalised with diarrhoea. METHODS: Immune serum was raised in rabbits by inoculating semipurified rotavirus, SA-11 strain. Immunoglobulins were conjugated to horse radish peroxidase and a rapid ELISA for rotavirus diagnosis was developed. The rapid ELISA was compared with routine ELISA, developed earlier at NIV. RESULTS: Of the 155 faecal samples from patients with diarrhoea, 96 were positive by rapid ELISA and 95 in routine NIV ELISA. OD values were higher in rapid ELISA. The rapid ELISA takes only 4 h to complete. INTERPRETATION & CONCLUSION: Rotavirus diagnosis by rapid ELISA is simple and easy to perform. This may lead to a significant reduction in the unnecessary usage of antibiotics, which cannot control infection due to rotavirus. This technology is being commercialized.
Subject(s)
Child , Diarrhea/diagnosis , Enzyme-Linked Immunosorbent Assay/methods , Humans , Rotavirus/isolation & purification , Rotavirus Infections/diagnosisABSTRACT
The prevalence of rotavirus diarrhea was compared in two settings, among children attending outpatient clinics and those hospitalized (inpatients) at Pune, India. A total of 489 and 628 fecal specimens were collected during October 1993 to September 1996 from outpatients and inpatients respectively. Overall occurrence of rotavirus diarrhea was more among hospitalized children. Using the stratification on the variable age, it is shown that age is indeed a confounding variable. The important finding of the study was, in < or = 6 months age group, it was observed that the occurrence of rotavirus diarrhea was more in the outpatients (30.26%) than among the inpatients (10.11%). Children of this age group are likely to be partially protected by maternal antibodies. The effect of seasonality and sex distribution did not differ in the two settings. It was found that G2 serotype was the major cause of diarrhea among the outpatients.
Subject(s)
Ambulatory Care Facilities , Diarrhea/epidemiology , Female , Hospitalization , Humans , India/epidemiology , Inpatients/statistics & numerical data , Male , Outpatients/statistics & numerical data , Prevalence , Rotavirus Infections/epidemiologyABSTRACT
Group A rotavirus-positive stool specimens, collected from 432 hospitalized patients of all age groups with diarrhoea during 1990-1997 from Pune, India, were characterized for subgroups (SGs) and G serotypes (1, 2, 3, 4, 6, 8, and 10). ELISA for subgrouping was carried out by employing subgroup I and II-specific monoclonal antibodies (MAbs). For serotyping, MAbs against G1 (Ku), G2 (S2), G3 (Yo), and G4 (ST-3) were used. In addition, MAbs against G3 (RV-3), G8 (B37), G6 (bovine U.K.), and G10 (B223) were also employed. Of the 432 specimens, 174 (40.27%) belonged to subgroup I, 187 (43.29%) to subgroup II, 15 (3.47%) to subgroup I and II, and 56 (12.96%) did not react to MAbs specific to subgroup I and subgroup II MAbs. Of the 432 specimens, 111 (25.69%) reacted to one of the MAbs used. Thirty-five of the 111 specimens were serotyped as G1, 34 as G2, and 42 as G3, G4, G6, G8, and G10. Sixty-seven (21%) specimens gave dual reaction mainly to MAbs against G6, G10; G2, and G4, and in several other combinations. Forty-seven specimens (10.88%) showed multireactivities. A large number of specimens (47.92%) did not show any reactivity with MAbs employed in this study, and remained non-serotypeable. Subgroup I was found to be more common in Pune, and most specimens negative for subgroup I and II were non-serotypeable. The results implicate the need for characterization of unusual and non-typeable strains before undertaking any rotaviral vaccine studies in India.
Subject(s)
Adolescent , Adult , Antibodies, Viral/analysis , Child , Child, Preschool , Diarrhea/epidemiology , Enzyme-Linked Immunosorbent Assay , Humans , India , Infant , Rotavirus/classification , Rotavirus Infections/epidemiology , Serotyping , VaccinationABSTRACT
Rotavirus was detected in 266 (28.15%) out of 945 faecal specimens collected between July 1992 and June 1996 from children < or = 5 yr of age. Statistical analysis using odds ratios and multivariate logistic regression analysis revealed that seasonality had a strong influence on the number of rotavirus diarrhoea cases admitted to the hospital. Maximum cases occurred in the winter and minimum in the rainy season. Age was strongly associated with the prevalence of rotavirus diarrhoea. The age group of 6-24 months was the most susceptible. This disease was more predominant in males.
Subject(s)
Adolescent , Adult , Age Distribution , Child , Child, Preschool , Diarrhea/epidemiology , Female , Hospitalization , Humans , India , Male , Prevalence , Rotavirus Infections/complications , Seasons , Sex DistributionABSTRACT
A total of 205 specimens positive for rotaviruses obtained during 1990-93 surveillance studies of hospitalized patients with diarrhoea were tested by ELISA for G serotypes. Of these, 107 (52.19%) specimens could be serotyped. Seventy four specimens were assigned to one of the G1-G4 serotypes. Of the 74 specimens, 49 (66.2%) reacted with MAb against G2. Of 107 typeable specimens, 33 showed dual or multiple reactivity. Eighteen specimens reacted with MAb specific to G1 and G2 serotypes. Five specimens reacted with MAb specific to G2 and G4. RNA profile of some of the specimens showing dual reactivities did not show additional RNA band. The specimens showing reaction to G1 and G2, had long RNA pattern, those with G2 and G4, showed short RNA pattern. Ten (9.17%) specimens reacted to 3 or more rotavirus serotypes. Of these, three reacted to MAb raised against G6, a bovine rotavirus. Ninety eight specimens could not be serotyped.
Subject(s)
Humans , India , Prevalence , Rotavirus/classification , Serotyping , Time FactorsABSTRACT
Preparation of reagents for the diagnosis of rotavirus by enzyme linked immunosorbent assay (ELISA) was carried out at the National Institute of Virology (NIV), Pune. This is a double antibody sandwich ELISA test. The coating antibody was raised in guinea pig against SA-11 (Simian rotavirus), and is used at 1:20,000 dilution. The indicator antibody was also raised against SA-11 in rabbits. The rabbit preimmune serum is used as negative control serum. Both, pre- and post-immune rabbit antisera are used at 1:10,000 dilution in the test. Goat IgG-HRP conjugate against rabbit IgG was procured commercially (Sigma, USA). Results of ELISA test can be read visually. A total of 63 pretested faecal specimens and 38 specimens of rotavirus in different concentrations were compared simultaneously by the NIV ELISA and Dakopatts ELISA. Of the 63 specimens, 36 were positive in NIV ELISA and 33 positive by Dakopatts ELISA. Human and animal rotavirus strains also showed higher titers in NIV ELISA than Dakopatts ELISA. From our data we conclude that NIV ELISA is 100 per cent specific and more sensitive than Dakopatts ELISA test. It is much more economical than any other commercial kit available for the diagnosis of rotavirus. Since the reagents are in lyophilized form, they are suitable for use in developing countries.