Comparative evaluation of antigen detection ELISA and reverse transcriptase PCR in acute stage of Japanese encephalitis prevalent in endemic areas of North-Eastern part of Uttar Pradesh, India (original) (raw)
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Transactions of the Royal Society of Tropical Medicine and Hygiene, 2012
Japanese encephalitis (JE), a neurotrophic disease, was first recorded in the state of West Bengal, India in 1973. Since then JE is being reported every year from different districts. With a view to identify the JE cases accurately, a study was undertaken to detect the Japanese encephalitis virus (JEV) as the etiologic agent from the acute encephalitis syndrome (AES) cases and to identify its distribution in different districts. We report the results of 513 blood or cerebrospinal fluid samples referred/collected from the hospitalized AES cases. The samples were initially subjected to Mac-ELISA test followed by reverse transcriptase (RT)-PCR for the detection of IgM antibodies and the JEV genome, specific to E gene, respectively. Out of 513 samples referred/collected, 139 (27.1%) samples were reactive to JE IgM antibody. The remaining 374 samples were screened to select those which had a history of illness with a duration of ≤3 days. Only 147 samples were selected and tested, out of which 36 (24.5%) isolates were achieved and those were RT-PCR positive against the control JEV strain. Detection of IgM antibody to JE and the RT-PCR result confirms the active circulation of JEV in different districts of West Bengal and needs to be monitored carefully.
Journal of Clinical Virology, 2005
Background: Japanese encephalitis (JE) is endemic in Cuddalore district, Tamil Nadu (TN), Southern India. The reports of JE cases from the local hospitals did not reflect the actual disease burden. It is likely that these cases were attending the nearby referral hospitals, for want of better treatment facilities. Objectives: Between July 2002 and February 2003, a pilot study was undertaken to examine whether JE was a component of paediatric acute encephalitis syndrome (AES) reported to two major referral hospitals adjacent to Cuddalore, and to map the distribution of the JE cases. Study design: A total of 58 hospitalized children [0-15 years] with AES were investigated. Other than the routine laboratory investigations, either CSF or sera or both [depending on the availability] collected from these children were analyzed at Center for Research in Medical Entomology, Madurai (TN) for JEV-antigen, antibody detection, virus isolation and virus genome detection by indirect immunofluorescence, MAC enzyme linked immunosorbent assay (ELISA), insect bioassay and by reverse transcriptase polymerase chain reaction (RT-PCR), respectively. Results: JE was established in 17 (29%) of 58 AES cases; half of the AES cases [31/58, 53%] and 59% [10/17] of JE cases were confined to JE-endemic areas in Cuddalore district. The JE confirmation scored by different assays varied according to the clinical phase of the illness. The attack rate was high among the children aged 3-8 years. The monthly distribution of acute encephalitic syndrome cases followed the distribution of JE cases [coinciding with the rainy season in this region] suggesting encephalitis of JE origin. Conclusion: In JE-endemic areas, the actual JE burden can be estimated by the collection of JE case reports from the local hospitals and from the referral hospitals. Building of diagnostic facilities in hospitals for JE is necessary to achieve this goal.
Journal of Clinical Virology, 2004
Background: Japanese encephalitis (JE) is endemic in Cuddalore district, Tamil Nadu (TN), Southern India. The reports of JE cases from the local hospitals did not reflect the actual disease burden. It is likely that these cases were attending the nearby referral hospitals, for want of better treatment facilities. Objectives: Between July 2002 and February 2003, a pilot study was undertaken to examine whether JE was a component of paediatric acute encephalitis syndrome (AES) reported to two major referral hospitals adjacent to Cuddalore, and to map the distribution of the JE cases. Study design: A total of 58 hospitalized children [0-15 years] with AES were investigated. Other than the routine laboratory investigations, either CSF or sera or both [depending on the availability] collected from these children were analyzed at Center for Research in Medical Entomology, Madurai (TN) for JEV-antigen, antibody detection, virus isolation and virus genome detection by indirect immunofluorescence, MAC enzyme linked immunosorbent assay (ELISA), insect bioassay and by reverse transcriptase polymerase chain reaction (RT-PCR), respectively. Results: JE was established in 17 (29%) of 58 AES cases; half of the AES cases [31/58, 53%] and 59% [10/17] of JE cases were confined to JE-endemic areas in Cuddalore district. The JE confirmation scored by different assays varied according to the clinical phase of the illness. The attack rate was high among the children aged 3-8 years. The monthly distribution of acute encephalitic syndrome cases followed the distribution of JE cases [coinciding with the rainy season in this region] suggesting encephalitis of JE origin. Conclusion: In JE-endemic areas, the actual JE burden can be estimated by the collection of JE case reports from the local hospitals and from the referral hospitals. Building of diagnostic facilities in hospitals for JE is necessary to achieve this goal.
Indian Journal of Microbiology Research
Japanese encephalitis Virus (JEV) a flaviviridae family member (genus Flavivirus) is the main cause of meta-zoonotic viral encephalitis in many Asian countries. The disease was primarily reported in 1952 from Nagpur territory of Maharashtra recording nearly 16 deaths of an unknown viral encephalitis which was later awarded to be JEV; just nearer to Chandrapur reporting the present catastrophes.Hence we have undertaken present study to make a countable move not only towards the diagnosis of the diseases but also to restrict its future spread in Indian continent. Which will also aid medical practitioners to combat with this metazoonotic disorder.Amongst total suspected population nearly these 20 males and 22 females were positive. Considering monthly distribution most number of suspected cases were seen in the month of July. Most number of JEV affected population was seen more in the age group of 1 to 5 years of children and with advancement of the age reduction in the number of serop...
Indonesian Journal of Tropical and Infectious Disease, 2013
Background: Japanese enchepalitis (JE) is a viral disease that considered as zoonotic disease, which transmitted through mosquito vectors that had JE virus. Mainly caused by the mosquito C. Tritaeniorhynchus (the most important vector is the mosquito Culex, which feeds on cattle in preference to human). JE virus disease can also cause disturbances in the central nervous system eg. brain, bone marrow, and meninges which has serious impact on public health. This disease has been reported from Japan, Korea, Taiwan, India, Myanmar, Thailand, Western Pacific and Southeast Asia to Indonesia. However, the incidence of this disease in Indonesia has not been well known in various animal species or humans. Aim: The purpose of this study is to develop rapid diagnostic examinations on patient diagnosed JE virus in Surabaya by using PCR (Polymerase Chain Reaction). Because, JE disease can lead to dead-end at the patient if not treated immediately. Method: The research methods, extraction method,...
The Journal of Infection in Developing Countries, 2009
Background: In the year 2005, an epidemic of Japanese encephalitis (JE) occurred in the northern states of India. The present study was planned to reconfirm the circulation of JE in the area and to assess the trend of the disease to slow down the burden of JE. Methodology: Surveillance was conducted to identify patients with acute encephalitis. Blood and cerebrospinal fluid specimens from suspected cases underwent pathological, serological, and demographic investigations. Viral testing for evidence of Japanese encephalitis virus (JEV) infection was also performed, either by IgM capture ELISA/RT-PCR or both. To identify circulating JEV strains, RT-PCR, sequencing and phylogenetic analysis was performed. Based on clinical cases reported between 1992 and 2008, the trend of JE infection in the state was analyzed to examine the dynamics of infection. Results: Our investigations (n = 38) revealed that only 55.3% cases were positive for JE. Pathological examination revealed marked pleocytosis in CSF (90 76.9 cells/mm 3 ), and peripheral leucocytosis (64.7 8.86% neutrophils) with mild anemia. Males were more susceptible than females with a ratio of 1.63:1 and significant gender difference (P 0.05) was observed in patients below six years. In the patient group younger than six years, the rate of infection per million was six-fold higher (P 0.005) in males as compared to females. Our phylogenetic study suggests that the circulating strain during the 2005 JE epidemic was close to GP78, and in the future a larger epidemic may occur. Conclusions: The 2005 JE epidemic was possibly caused by JEV GP78 and it is spreading into newer areas. The trend of JE suggests that the problem in North India is escalating and larger epidemics may occur in the future; therefore, serious steps are necessary to combat JE, including the development of more efficient surveillance methods and differential diagnosis.
Journal of Medical Virology, 2004
A rapid, sensitive, and accurate laboratory diagnostic test is needed for distinguishing Japanese encephalitis virus (JEV) from other diseases featuring similar clinical symptoms and also for preventing potential outbreaks. In this study, a TaqMan reverse transcription (RT)-polymerase chain reaction (PCR) assay was developed for rapid detection and quantification of the viral RNA of various JEV strains. A consensus JEV NS3 region was chosen to design the primers and the TaqMan probe. The JEV TaqMan assay used the EZ-rTtH RT-PCR system featuring advantages such as a one-step, high-temperature RT reaction modality and preventing carry-over contamination. The sensitivity of the JEV TaqMan assay for detecting in vitro-transcribed JEV NS3 RNA was estimated to be one to five copies of RNA per reaction. For cultured JE virions, less than 40 plaque forming unit (PFU)/ml of virus load (corresponding to 0.07 PFU/test) could be detected. In addition, the JEV TaqMan assay could detect all seven strains of JEV tested, but provided negative results for nine other flaviviruses and encephalitis viruses tested. The JEV TaqMan assay demonstrated greater sensitivity and specificity than traditional RT-PCR methods as has been previously reported. The application of the JEV TaqMan assay herein has been shown to the sensitive detection of the JEV from both mosquito pools and also JEV-spiking human blood. The assay should be of use in diagnostic laboratory conduct and could be used to replace or complement time-consuming viral-culture methods, thus achieving more rapid, sensitive, and highly specific identification of JEV infection.