Comparison of JEV neutralization assay using pseudotyped JEV with the conventional plaque-reduction neutralization test

Abstract

We previously reported the development of a neutralization assay system for evaluating Japanese Encephalitis Virus (JEV) neutralizing antibody (NAb) using pseudotyped-JEV (JEV-PV). JEV-PV-based neutralization assay offers several advantages compared with the current standard plaque-reduction neutralization test (PRNT), including simplicity, safety, and speed. To evaluate the suitability of the JEV-PV assay as new replacement neutralization assay, we compared its repeatability, reproducibility, specificity, and correlated its results with those obtained using the PRNT. These analyses showed a close correlation between the results obtained with the JEV-PV assay and the PRNT, using the 50% plaque reduction method as a standard for measuring NAb titers to JEV. The validation results met all analytical acceptance criteria. These results suggest that the JEV-PV assay could serve as a safe and simple method for measuring NAb titer against JEV and could be used as an alternative approach for assaying the potency of JEV neutralization.

MLKL Mediated Necroptosis Accelerates JEV-Induced Neuroinflammation in Mice

“…Next, the supernatant was ultra-centrifuged at 40,000 g for 4 h at 4°C and the viral pellet was resuspended with PBS at 100:1 (supernatant: suspension) after removing the supernatant thoroughly. After being filtered through 0.22 μm filters, the virus suspension was titrated by conventional plaque assay and stored at −80°C (Lee et al, 2014 ).…”

A Novel Silver Nanoparticles-based Sensing Probe for the Detection of Japanese Encephalitis Virus Antigen

“…Several conventional diagnostic methods have been developed for the detection of JEV infection, such as, enzyme-linked immunoglobulin assay (ELISA), plaque reduction neutralization test (PRNT) (Lee et al 2014;Sirivichayakul et al 2014) and reverse transcription polymerase chain reaction (RT-PCR) (Swami et al 2008). Nonetheless, all of these diagnostic techniques have disadvantages such as high cost of consumables, expensive instrumentation and long analysis assay time .…”

Development of a neutralization assay based on the pseudotyped chikungunya virus of a Korean isolate

“…In addition, it takes a relatively long time (about 4-5 days) to visualize and count the plaques for the PRNT. To overcome these limitations, alternative neutralization assays using non-infectious virus replicon particles (VRPs) or pseudotyped viruses have been developed (Gläsker et al, 2013;Kishishita et al, 2013;Hu et al, 2014;Lee et al, 2014). Here, we adopted the strategy of expressing the entire structural polyprotein of CHIKV to generate the pseudotyped virus form of KNIH/2009/77 (CHIKVpseudo) for the enhanced infectivity compared to a virion expressing either an individual E1 or E2 protein (Hu et al, 2014).…”