Prevalence of West Nile virus infection in India.

During the course of the virological investigation of cases of suspected viral fevers carried out at the National Institute of Virology (NIV), Pune, India, evidence of recent infection with West Nile (WN) virus was detected in 88 cases. Fever, general aches, headache, nausea and vomiting were the principal clinical features in 92% (81/88) of the cases; there were seven cases of encephalitis, in which WN virus-specific IgM class antibodies were detected in CSF samples. These cases of encephalitis were from Japanese encephalitis (JE) nonendemic areas, like Maharashtra and Rajasthan, as well as from JE endemic areas, like Goa and Orissa. Interestingly, neutralizing antibodies predominantly to WN virus were detected in CSF samples by the 50% cytopathic effect inhibition method; the titers ranged from 5 to 375. Cases of WN virus infection associated with both encephalitis and classic features have been reported for the first time in recent years in India. Reports of unique urban West Nile virus encephalitis epidemics in New York, Romania, and Algeria in recent years have signaled the emergence of neurological infection due to West Nile virus as a novel public health threat. This study is important because it records evidence of WN virus infection in India.

Development of monoclonal antibody based antigen capture ELISA to detect chikungunya virus antigen in mosquitoes.

BACKGROUND & OBJECTIVES: Chikungunya (CHIK) virus has caused numerous large outbreaks in India. No active or passive surveillance has been carried out since the last epidemic which occurred in 1971. For active surveillance, it is necessary to have a test, which can detect the virus from a large number of field-collected mosquitoes. METHODS: The present study describes the standardization of monoclonal antibody (MAb) based antigen capture ELISA to detect chikungunya virus antigen from the mosquitoes. CHIK virus antigen from suspension of experimentally infected mosquitoes and their progeny was captured on mouse polyclonal antibody, while biotinylated CHIK Mab was used as a probing antibody. CHIK virus antigen in the head squashes of virus inoculated mosquitoes was detected using indirect immunofluorescence antibody (IFA) test for confirmation of ELISA results. RESULTS: The ELISA test was sensitive enough to detect antigen even if a small fraction of a single infected mosquito homogenate was incorporated in the test. The IFA test failed to detect CHIK antigen in 10 and 25 microliters of suspension whereas with ELISA it was detected in all the samples. Progeny of Aedes aegypti and Ae. albopictus mosquitoes infected with chikungunya virus did not show the possibility of existence of transovarial transmission. INTERPRETATION & CONCLUSION: This test is rapid and simple since it can be completed in two days as compared to the conventional mosquito inoculation and IFA techniques, which require at least 10 days. There is an additional advantage with this test that a large number of samples can be processed, and the remaining homogenate of the mosquitoes can be used for screening other viruses. Experimental data raised using this test showed that transovarial transmission of this virus does not occur in these vector species.

An outbreak of dengue fever in rural areas of northern India.

During the past few decades, epidemics of dengue fever are causing concern in several South-East Asian countries including India. The rural areas of Hissar district of Haryana state, situated about 170 km North-West of Delhi, experienced an outbreak of febrile illness during July-August 1996. A total of 13 villages in eight affected primary health centres reported fever cases. The clinical, epidemiological and entomological findings indicated that the present episode was due to dengue fever. The aetiological agent of the current outbreak, the DEN-2 virus, was isolated from 12 acute-phase sera specimens. Though, in the recent past outbreaks have been reported from the rural areas of southern and western India, the present episode is the first outbreak being reported from the rural areas of northern India. The increasing frequency of dengue fever outbreaks in rural areas of various Indian states reflects the changing life style of the rural population as a result of urbanization process and calls for a suitable prevention and control policy based on strengthened surveillance, appropriate health education to the community coupled with proper training of health personnel.

Detection of Coxsackievirus B 4 antibodies in cases of suspected encephalopathy by microneutralization test.

A retrospective study of serum and cerebrospinal fluid (CSF) samples collected from suspected viral encephalitis and encephalopathy cases was carried-out and it included 100 CSF and 89 serum samples from Goa, collected during 1990-1994. These samples which were negative for antibodies to Japanese encephalitis (JE), West Nile (WN), Dengue-2 (DN-2) and herpes viruses, were tested for Coxsackievirus B 4 specific antibodies by 'in vitro' microneutralization technique along with 80 negative control serum samples. Out of 189 specimens (100 CSF and 89 serum), 23 CSF and 41 serum samples were positive for Coxsackievirus B 4 neutralizing antibodies. Antibody profile seemed to be IgG as revealed by mercaptoethanol treatment. The presence of neutralizing antibodies to coxsackievirus B 4 with titres as high as 1:512 in 8 CSF and 19 serum samples seemed to be suggestive of viral meningitis due to Cox B-4 viruses.

Japanese encephalitis in Sangli district, Maharashtra.

During the months June to December, 1997, 52 cases of suspected viral encephalitis were admitted at the Government Hospital, Sangli. These cases were from the congested areas of Sangli and the adjoining villages. All age groups and both genders were affected. IgM antibodies to Japanese encephalitis (JE) virus were detected in the sera of five of the 52 cases. Perhaps JE virus has established itself in a new locality in Maharashtra and could cause serious public health problems.

Hemorrhagic manifestations and encephalopathy in cases of dengue in India.

Thirty-seven serum samples and five serum-CSF pairs collected from 42 acutely ill patients admitted to hospitals in Maharashtra (Bombay, Pune and Nasik); Orissa (Raurkela) and South Goa were referred to the National Institute of Virology (NIV), Pune (Maharashtra, India) for serodiagnosis. These patients had clinical manifestations of fever, hemorrhagic manifestations, hepatomegaly, shock syndrome and encephalopathy. Sixty-six percent of patients were children below ten years of age. Serological investigations revealed infection to dengue virus in all the patients as indicated in detection of IgM antibodies predominantly to dengue viral antigens. An important outcome of the study is that 10 patients referred to NIV with a provisional diagnosis of viral encephalitis proved to be dengue.

The 1993 epidemic of dengue fever in Mangalore, Karnataka state, India.

An epidemic of febrile illness with hemorrhagic manifestations occurred in certain parts of Mangalore city, Karnataka state, India, from the last week of July 1993. The epidemic reached its peak by mid-August and then started declining. Sporadic cases, however, continued to occur till early December. About 200 cases were reported covering all age groups and both sexes. The cases presented with pyrexia, myalgia, arthralgia and headache. Palatal petechiae, magenta colored tongue with central coating, maculopapular rash and facial flush were observed as classical signs. The tourniquet test was positive in 12% of the cases. Hemorrhage was observed in the form of epistaxis (2 cases), subconjunctival hemorrhage (2 cases) or purpura (3 cases). There were no deaths which were attributable to the epidemic. Five strains of dengue (DEN-2) virus were recovered from the acute-phase sera. Dengue virus-specific IgM type of antibodies were detected in 29/116 (25%) sera. Breeding of Aedes aegypti was observed in some of the areas where cases had occurred. No virus was isolated from any of the field-caught Ae. aegypti mosquitos.

Detection of virus specific IgG subclasses in Japanese encephalitis patients.

During the Japanese encephalitis (JE) epidemic in 1988 at Gorakhpur, Uttar Pradesh, 34 cerebrospinal fluid (CSF) samples with 16 matching sera from 34 anti JEV IgM positive (confirmed JE) and 24 CSF samples with 4 matching sera from 24 anti JEV IgM negative (clinical encephalitis) patients were collected and tested for presence of JEV specific IgG by ELISA. Eighteen CSF samples and 8 matching sera from confirmed JE and 5 CSF samples and one matching serum from clinical encephalitis patients positive for JEV specific IgG were further assayed for subclass specificity using specific murine monoclonal antibodies. Almost all the samples exhibited IgG1 as the virus specific subclass. In addition to IgG1, one serum and one CSF sample each from two different confirmed JE patients showed the presence of virus specific IgG4 and IgG3 respectively. Half of the confirmed JE and clinical encephalitis patients exhibited intrathecal synthesis as evident from either elevated IgG index or CSF IgG/CSF albumin ratio. Most of the patients who recovered had predominantly virus specific IgG1 in CSF. It seems likely that IgG1 might have a protective role in clearance of virus from the central nervous system.