Comparison of JEV Neutralization Assay Using Pseudotyped JEV with the Conventional Plaque-Reduction Neutralization Test

Hee-Jung Lee; Kyung-Il Min; Ki Hoon Park; Hyo Jung Choi; Min-Kyoung Kim; Chi-Young Ahn; Young-Jin Hong; Young Bong Kim

doi:10.1007/s12275-014-3529-y

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Validation Study
Comparison of JEV Neutralization Assay Using Pseudotyped JEV with the Conventional Plaque-Reduction Neutralization Test: Hee-Jung Lee ¹, Kyung-Il Min ², Ki Hoon Park ¹, Hyo Jung Choi ¹, Min-Kyoung Kim ¹, Chi-Young Ahn ³, Young-Jin Hong ⁴, Young Bong Kim ¹; Journal of Microbiology 2014;52(5):435-440.
DOI: https://doi.org/10.1007/s12275-014-3529-y
Published online: March 7, 2014

¹Department of Bio-industrial technologies, Konkuk University, Seoul 143-701, Republic of Korea, ²Department of Biopharmaceuticals and Herbal Medicine Evaluation, National Institute of Food and Drug Safety Evaluation, Osong 363-951, Republic of Korea, ³Blood Products Team, National Institute of Food and Drug Safety Evaluation, Osong 363-951, Republic of Korea, ⁴Department of Pediatrics, College of medicine, Inha University, Incheon 402-751, Republic of Korea

Corresponding author: Young Bong Kim , Tel: +82-2-450-4208,

Received: 10 October 2013 • Revised: 13 December 2013 • Accepted: 16 December 2013

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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 advan-tages compared with the current standard plaque-reduc-tion 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 re-duction 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.

Keywords: Japanese encephalitis virus;neutralizing antibody;pseudovirus;PRNT

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Comparison of JEV Neutralization Assay Using Pseudotyped JEV with the Conventional Plaque-Reduction Neutralization Test

J. Microbiol. 2014;52(5):435-440. Published online March 7, 2014

DOI: https://doi.org/10.1007/s12275-014-3529-y

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