Rapid ELISA for the diagnosis of rotavirus.

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.

A serological survey of arboviral diseases among the human population of the Andaman and Nicobar Islands, India.

In an attempt to determine the prevalence of certain arthropod-borne viruses of public health importance amongst the human population of the Andaman and Nicobar Islands of India, 2,401 sera were collected from six major localities. The sera were analysed by the hemagglutination inhibition (HI) and neutralization (N) tests, using Chikungunya (CHIK), Japanese encephalitis (JE), West Nile (WN), dengue (DEN-2), Langat (TP-21) and Kyasanur Forest disease (KFD) viral antigens. The highest prevalence of HI antibodies was detected against KFD virus (22.4%), followed by Langat (20.2%), JE (5.9%), DEN-2 (3.1%), CHIK (2.9%) and WN (0.8%) viruses. Cross-reactions to the viral antigens were also noted. The results of N tests indicated a high prevalence of DEN-2 (25.4%) virus, followed by Langat (17.5%), CHIK (15.3%), KFD (12%), JE (2.19%) and WN (1.8%). These results are discussed in relation to important epidemiological parameters like age, sex and geographical location. To our knowledge, this is the first report of an extensive serosurvey of arthropod-borne viruses on these islands.

In vitro antiviral activity of bael (Aegle marmelos Corr) upon human coxsackieviruses B1-B6.

The in-vitro antiviral activity of a series of compounds in samples extracted from various parts of the Indian holy tree, Bael (Aegle marmelos corr.) were evaluated for their efficacy against human coxsackieviruses B1-B6. The inhibitory concentrations (IC50) for leaves (L1 and L2) stem and stem bark (S1, S2, S3 and S4) fruit (F1 and F2micro) root and root bark (R1 and R2) and pure compound, the marmelide were 1000 microg/ml (for L1 and L2), 1000 microg/ml (for S1, S2, S3 and S4), 1000 microg/ml (for F1) and 500 microg/ml (for F2) 250 microg/ml (for R1) and 500 microg/ml (for R2) and 62.5 microg/ml for marmelide respectively by plaque inhibition assay at 96 hrs. On the other hand, the corresponding value for Ribavirin, a standard antiviral drug, was 2000 microg/ml for the same viruses at the same time period. These concentrations did not exhibit any toxicity to Vero cells, the host subtoxic concentrations were 5000 microg/ml for leaf and stem fractions 2000 microg/ml for fruit fractions 500 and 1000 microg/ml for root fractions 250 microg/ml for marmelide and 2000 microg/ml for Ribavirin. The cytotoxic concentrations were 8000 microg/ml for leaf and stem compounds 4000 mg/ml for fruit; 1000 microg/ml and 2000 microg/ml for root 500 microg/ml for marmelide and 4000 microg/ml for ribavirin at 96 hrs. These were also confirmed by trypan blue dye exclusion test and further passaging of cells. Additionally pretreatment of host cells, virus inactivation, yield reduction and effect of time of addition assays against coxsackievirus B3 suggested that marmelide was most effective as a virucidal agent besides interfering at early events of its replicative cycle like adsorption, penetration, at various steps in single cycle growth curve and effect of time of addition.

'In vitro' antiviral activity of neem (Azadirachta indica. A. Juss) leaf extract against group B coxsackieviruses.

The antiviral and virucidal effect of methanolic extract fraction of leaves of neem (Azadirachta indica A. Juss) (NCL-11) was studied regarding its activity and possible mechanism of action against Coxsackie B group of viruses. NCL-11 inhibited plaque formation in 6 antigenic types of Coxsackie virus B at a concentration of 1000 micrograms/ml at 96 hrs. 'in vitro'. Additionally virus inactivation, yield reduction and effect of time of addition assays suggested that NCL-11 was most effective against coxsackie virus B-4 as a virucidal agent besides interfering at an early event of its replicative cycle. The evidence suggested that presence of a battery of compounds besides flavonoids, triterpenoids and their glycosides in NCL-11 have antiviral action for coxsackie B group of viruses 'in vitro.' The minimal inhibitory concentrations were not toxic to Vero (African green monkey kidney), cells; subtoxic concentration was 8,000 micrograms/ml and cytotoxic concentration 10,000 micrograms/ml, which was confirmed by trypan blue dye exclusion test.

Comparative efficacy of three measles vaccines in Indian children.

A Comparative study of three types of measles vaccines was undertaken among 1005 children. Of these 527 were vaccinated with the Serum Institute of India (SII) vaccine, 230 with Schwarz (SC) and 248 were vaccinated with Edmonston-Zegreb (EZ) vaccine (imported from Zegreb). Though the majority of children reacted favourably with all the three vaccines (SII: 98.43%; SC: 93.40%; EZ: 93.0%) with a rise in titre, but the percentage of seroconversion was significantly higher with the SII vaccine (p < 0.01). The Schwarz and Edmonston Zagreb vaccines showed significantly less GM titre as compared with the other age group i.e. 9-12 months (p < 0.05). With Serum Institute of India (SII) vaccine the GM titres were almost similar in the different age groups. The overall GM titre obtained with the SII vaccine was significantly higher than the SC vaccine (p > 0.001) as well as the EZ vaccine (p > 0.001). It is of interest to note that among the infants, 22.5% children had measles antibody in them before vaccination.

Activity of dengue-2 virus and prevalence of Aedes aegypti in the Chirimiri colliery area, Madhya Pradesh, India.

Repeated outbreaks of a suspected viral fever in Chirimiri colliery area, Madhya Pradesh were reported since 1990. The area consists of an agglomeration of sprawling settlements at varying altitudes of 816 to 890 m and it has partial sylvan cover. During a 1992 outbreak 25 patients' sera were tested, of which 13 showed seropositivity to dengue (DEN) by MAC-ELISA test; DEN-2 was isolated from Aedes aegypti collected from two of the eight settlements of the area. The principal vector, Ae. aegypti, was prevalent in all the settlements studied; Breteau indices (BI) varied between 2.5 and 125.0; adult house indices (AHI) between 0 and 60.0%; Ae. albopictus and Ae. vittatus occurred in considerable numbers; Ae. aegypti bred in more containers with nonpotable water than those with potable water; the breeding of this species was noted in a maximum number of cement tanks while mud pots were predominant among the available containers. Paired comparisons between relative prevalence indices showed significant correlation and regression coefficients. Significant association of Ae. aegypti breeding with the households having tap water supply was noted, the relative risk declining with the people's use of well water either exclusively or in combination with other sources of water supply. It was also collected in the nonresidential areas. The role of ecological factors in the maintenance and spread of Ae. aegypti and dengue in these settlements is discussed.

Measles outbreak in a tribal population of Thane district, Maharashtra.

In March 1992, an outbreak of measles, in the tribal population of Vavar village, Mokhada Taluk, Thane district, Maharashtra, was investigated. Two hamlets of Vavar village namely Sagpanipada (epidemic in October, November 1991) and Behedpada (epidemic in January, February 1992) were affected. In both hamlets, measles cases were confined to children below 10 yrs and 96% of the cases occurred in children below 6 yrs. Attack rates were 52.7% and 51.4% and case fatality rates were 31.2% and 15.6% at Sagpanipada and Behedpada, respectively. All the convalescent patients' sera possessed IgM antibodies against measles. A clear drop in IgM and a rise in IgG antibodies against measles was observed in 35 paired samples from convalescent patients. Fifty four per cent of sera from controls, possessed IgM antibodies. Migrating population appeared to have imported measles which flared up in an epidemic among the susceptibles. Priority immunization of the children of remote isolated populations may prevent such epidemics.

Experience with a measles vaccine manufactured in India.

Five hundred and twenty seven children between 7 months and 2 years of age were vaccinated with measles vaccine manufactured by the Serum Institute of India. The sero-conversion rate in children who had no antibodies previous to vaccination was 98.4% as tested in HI. Ninety per cent of children who had pre-vaccination measles antibodies showed a two-fold or more rise in HI antibodies. The side reactions of the vaccine were negligible.