Japanese encephalitis is one of the serious vector-borne viral encephalitis diseases found worldwide and poses a major threat to public health. Most Japanese encephalitis virus (JEV) infections are subclinical; only 1: 250 to 1:1000 infected persons develop clinical presentations. Delay in proper diagnosis of JE affects the timeliness of treatment initiation and increases the mortality rate in patients. Therefore, there is an extreme need to develop potential biomarkers, which might improve the diagnosis and can become the basis for development of new therapeutics. The microRNAs (miRNAs/or miRs) are small noncoding RNAs of 17-24 nucleotides that are known to regulate about 60% of human genes. Although miRNAs have been found to regulate various aspects of innate and adaptive immune responses, less information on circulating miRNAs in JE is known. The study of JEV infected human serum miRNAs will provide novel information for the diagnosis of JE as well as for the improvement of disease outcome.Total RNA, including miRNA, was extracted from serum followed by the complementary DNA (cDNA) synthesis by using sequence-specific primers. cDNA was amplified using target-specific TaqMan MicroRNA Assay. Real-time polymerase chain reaction data was normalized using both exogenous (cel-miR-39) and endogenous (hsa-miR-93) controls.We have found significantly altered expression of miR-155 and miR-21 in serum of JEV infected patients as compared to healthy controls, revealing their role as a a noninvasive biomarker in JE. A significant correlation between miRNAs and JE was observed that offers the basis for miRNAs to serve as a new component to develop possible therapeutic strategies for JE in near future.
Viral infections of the central nervous system (CNS) occur sporadically and have been extensively studied because of the potential for permanent neurological damage or death. The neurotropic viruses have been reported to lead to various CNS infections. The objective of the present study is to develop an antigen detection ELISA protocol for detection and quantification of viral antigen in CNS infections by assessing the usefulness of antipeptide antibodies against potential peptides of cytomegalovirus (CMV), Epstein-Barr virus (EBV), varicella zoster virus (VZV), Japanese encephalitis virus (JEV), dengue (DENV), West Nile virus (WNV) and Chandipura virus (CHPV). A total of 182 cerebrospinal fluid (CSF) samples from confirmed, suspected and non-viral infections of the CNS were evaluated using panels of antipeptide antibodies against synthetic peptides of viral proteins. The cases of confirmed and suspected viral infections of the CNS showed 77% and 11% positivity, respectively, for the detection of viral antigen using antipeptide against synthetic peptides of CMV, EBV, VZV and JEV. The concentration of viral antigen was also obtained by using antipeptide of respective viruses in CSF from both the groups. The viral antigen concentration was also correlated with viral load in confirmed cases of viral infection of the CNS. This study demonstrates the use of antipeptide against synthetic peptide derived from CMV, EBV, VZV and JEV in diagnostics of viral infections of the CNS using patients' CSF samples.
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