Detection and Isolation of Japanese Encephalitis Virus From Blood Clots Collected During the Acute Phase of Infection

Abstract

Clinical specimens from an encephalitis outbreak in the Lakhimpur area of Uttar Pradesh, India, were investigated for identification and characterization of the etiologic agent. IgM capture ELISA showed recent Japanese encephalitis virus (JEV) infection. JEV isolation was attempted from white blood cells (WBCs) separated from blood clots of 12 patients (9 IgM positive and 3 negative) by serial co-culturing with phytohemagglutinin P-stimulated peripheral blood mononuclear leukocytes (PBMCs) obtained from pre-screened JEV sero-negative healthy individuals. JEV was isolated from two IgM-positive blood clots. Isolate 014178 was detected in WBCs and in the first passage of PBMCs by ELISA and reverse transcriptase-polymerase chain reaction. Isolate 014173 was detectable only after a second passage in PBMC co-culture. Sequence analysis of 346 nt of the C-prM region showed homology with JEV strain GP78. This is the first report on isolation of JEV from patient blood clots. Our study shows that the co-cultures of PBMCs separated from patient blood clots provide an additional source for JEV isolation.

miR-146a suppresses cellular immune response during Japanese encephalitis virus JaOArS982 strain infection in human microglial cells

“…JEV maintains a zoonotic life cycle where pigs are major reservoir hosts and mosquitoes act as vectors [3]. JEV infects macrophages and PBMCs [4] and infected macrophages help the virus to cross blood brain barrier [5]. However JEV also persists latently in T-lymphocytes [6] and PBMCs [7].…”

Acute Encephalitis Syndrome in Adults and Its Correlation with Cytokine Levels in the Serum and Cerebrospinal Fluid

“…A real-time PCR assay for quantification of JE viral RNA was also performed from serum and CSF samples of all patients, according to procedures described by Sapkal et al (11). Methods were developed by National Institute of Virology Pune India indigenously.…”

Acute Encephalitis with Atypical Presentation of Rubella in Family Cluster, India

“…We tested serum and urine samples from the index case-patient and serum, urine, and cerebrospinal fluid (CSF) samples from the third case-patient for Japanese encephalitis virus ( 5 ), West Nile virus ( 6 ), Chandipura virus ( 7 ), and enteroviruses ( 8 ) using real-time quantitative reverse transcription PCR (qRT-PCR); for dengue, chikungunya, and Zika viruses using CDC Trioplex qRT-PCR; and for rubella using RT-PCR ( 9 ). We further used quantitative PCR to test for varicella zoster virus DNA ( 10 ) and PCR for herpes simplex virus 1 (Genekam Biotechnology, Duisburg, Germany, No.…”