DOI: http://dx.doi.org/10.18203/2320-6012.ijrms20151166

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

Rajesh Kumar Verma, Dharmendra Prasad Singh, Ramakant Yadav, Ramakant Rawat

Abstract


Background: Objective of current study was to compare and evaluate different diagnostic tests to establish a reliable and less time consuming diagnostic test for prompt diagnosis of acute Japanese encephalitis cases mainly amongst children and young adults from North-Eastern part of Uttar Pradesh.  

Methods: A total of 100 subjects, including 50 suspected cases and 50 diseased controls were investigated to establish the diagnosis of JE in acute encephalitis patients. All CSF samples were subjected to MAC-ELISA, virus cultivation and RT-PCR.

Results: Out of 50 cases, 6 showed negative results by all the three tests. 50% (22 out of 44), 47.5% (21 out of 44), and 66% (23 out of 44) were found positive by MAC-ELISA, Virus cultivation and RT-PCR respectively. The rate of detection by Virus cultivation and RT-PCR was higher in case of MAC-ELISA negative cases (57.1% and 78.5%) than that of MAC-ELISA positive cases (22.7% and 27.2%) respectively. Mean age of the patients was 12.5 years, which ranged from 1-24 years with male to female ratio of 3:1.

Conclusions: The RT-PCR was found most reliable, sensitive and specific method amongst the three chosen methods for detection of JEV in suspected encephalitis patients.


Keywords


Antigen detection ELISA, Japanese encephalitis, MAC-ELISA, RT-PCR

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References


Okuno T. An epidemiological review of Japanese encephalitis. World Health Stats Rep. 1978;31:120.

Umenai T, Krzysko R, Bektimirov TA, Assaad FA. Japanese encephalitis: current worldwide status. Bull World Health Organ. 1985;63(4):625.

Medappa N. Japanese encephalitis in India. ICMR Bull. 1980;10:29.

Shope R. Medical significance of togaviruses: an overview of diseases in man and in domestic and wild vertebrate animals. In: Schlesinger RW, eds. The Togaviruses. 1st ed. New York: Academic Press; 1980: 47-82.

Monath TP. Pathobiology of flaviviruses. In: Schlesinger S, Schlesinger M, eds. The Togaviruses and Flaviviruses. 2nd ed. New York: Plenum Press; 1986.

Sabchareon A, Yoksan S. Japanese encephalitis. Ann Trop Pediatr. 1998;18:67.

Hoke CH, Nisalak A, Sangawhipa N, Jatanasen S, Laorakapongse T, Innis BL, et al. Protection against Japanese encephalitis by inactivated vaccines. N Engl J Med, 1988;319:608.

Rodrigues JJ, Shaikh BH, Shendarkar SP, Ramdasi SG. Laboratory confirmed cases of Japanese encephalitis in Maharastra state. Indian J Med Res. 1980;72:471- 4.

Work TK, Shah KV. Serological diagnosis of Japanese B type of encephalitis in North Arcot district of Madras state, India with epidemiological notes. Indian J Med Sci. 1956;10:582.

Narasimham MVVL, Krishna Rao C, Bendle MS, Yadava RL, Johri YC, Pandey RS. Epidemiological investigations on Japanese encephalitis outbreak in U.P. during 1988. J Comm Dis. 1988;20:263.

Kumar R, Mathur A, Kumar A, Sharma S, Saksena PN, Chaturvedi UC. Japanese encephalitis - an important cause of acute childhood encephalopathy in Lucknow, India. Postgrad Med J. 1988;64:18.

Mathur A, Tandon HO, Mathur KR, Sarkari NB, Singh UK, Chaturvedi UC. Japanese encephalitis virus infection during pregnancy. Indian J Med Res. 1985;81:9.

Kumar R, Mathur A, Kumar A, Sharma S, Chakraborty S, Chaturvedi UC. Clinical features and prognostic indicators of Japanese encephalitis in children in Lucknow (India). Indian J Med Res. 1990;91:321.

Dhillon GP, Raina VK. Epidemilogy of Japanese encephalitis in context with Indian scenario. J Indian Med Assoc. 2008;106(10):660-3.

Chatterjee S, Chattopadhyay D, Bhattacharya MK, Mukherjee B. Serosurveillance for Japanese encephalitis in children in several districts of West Bengal, India. Acta Paediatr. 2004;93:390-3.

Kabilan L, Vrati S, Rameshd S, Srinivasan S, Appaiahgari MB, Arunachalam N, et al. Japanese encephalitis virus (JEV) is an important cause of encephalitis among children in Cuddalore district, Tamil Nadu, India. J Clin Virol. 2004;31:153-9.

Chambers TJ, Hahn CS, Galler R, Rice CM. Flavivirus genome organization, expression and replication. Annu Rev Microbiol. 1990;44:649-88.

Smiyoshi H, Mori C, Fuke I, Morita K, Kuhara S, Kondou J, et al. Complete nucleotide sequence of the Japanese encephalitis virus genome RNA. Virology. 1987;161:497-10.

Mackenzie JS, Barrett ADT, Deubel V. Japanese encephalitis and West Nile viruses. Heidelberg, Germany: Springer; 2002: 12, 19, 36, 40.

Calisher CH, Beaty BJ. Laboratory diagnosis of viral infections. In: Lennette EH, eds. 2nd ed. New York: Marcel Dekker, Inc.; 1992.

Kumar R, Mathur A, Singh KB, Sitholey P, Prasad M, Shukla R, et al. Clinical sequelae of Japanese encephalitis in children. Indian J Med Res. 1993;97:9.

Bharadwaj M, Khanna N, Mathur A, Chaturvedi UC. Effect of macrophage derived factor on hypoferraemia induced by Japanese encephalitis virus in mice. Clin Exp Immunol. 1991;83:215.

Ehrenkranz NJ, Zemel ES, Bernstein C, Slater K. Immunoglobulin M in the cerebrospinal fluid of patients with arbovirus encephalitis and other infections of the central nervous system. Neurology. 1974;24:978-80.

Solomon T, Thao LTT, Dung NM, Kneen R, Hung NT, Nisalak A, et al. Rapid diagnosis of Japanese encephalitis by using an IgM dot enzyme immunoassay. J Clin Microbiol. 1998;36(7):2030.

Mathur A, Kumar A, Sharma S, Kulshreshtha R, Kumar R, Chaturvedi UC. Rapid diagnosis of Japanese encephalitis by immunofluorescent examination of cerebrospinal fluid. Indian J Med Res. 1990;91:1.

Burke DS, Lorsomrudee W, Leake CJ, Hoke CH, Nisalak A, Chongswasdi V, et al. Fatal outcome in Japanese encephalitis. Am J Trop Med Hyg. 1985;34:1203.

Ravi V, Vanajakshi S, Gowda A, Chandramuki A. Laboratory diagnosis of Japanese encephalitis using monoclonal antibodies and correlation of findings with outcome. J Med Virol. 1989b;29:221.

Gajanana A, Samuel P. Philip, Thenmozhi V, Ranendran R. An appraisal of some recent diagnostic assays for Japanese encephalitis. Southeast Asian J Trop Med Public Health. 1996;27:673.

Leake CJ, Burke DS, Nisalak A, Hoke CH. Isolation of Japanese encephalitis virus from clinical specimens using a continuous mosquito cell line. Am J Trop Med Hyg. 1986;35:1045-50.

Desai A, Ravi V, Guru SC, Slankar SK, Kaliaperumal VG, Chandramuki A, et al. Detection of autoantibodies to neural antigens in the CSF of Japanese encephalitis patients and correlation of findings with the outcome. J Neurol Sci. 1994;122:109.

Puri EA. Henchal, J. Burans, K. R. Porter, W. Nelson, D. M. Watts, C. G. Hayes. A rapid method for detection and identification of flaviviruses by polymerase chain reaction and nucleic acid hybridization. Arch Virol. 1994;134:29-37.

Fang Meiyu, Chen Huosheng, Chen Cuihua, Tian Xiaodong, Jiang Lianhua, Peng Yifei, et al. Detection of flaviviruses by reverse transcriptase polymerase chain reaction with the universal primer set. Microbiol Immunol. 1997;41:209-13.