Establishment of a Multiplex RT-PCR for the Sensitive and Differential Detection of Japanese Encephalitis Virus Genotype 1 and 3.
10.4167/jbv.2016.46.4.231
- Author:
Dong Kun YANG
1
;
Ha Hyun KIM
;
Hyun Ye JO
;
Sung Suk CHOI
;
In Soo CHO
Author Information
1. Viral Disease Division, Animal and Plant Quarantine Agency, Gyeongsangbuk-do, Korea. yangdk@korea.kr
- Publication Type:Original Article
- Keywords:
Japanese encephalitis virus;
Multiplex RT-PCR;
Genotype
- MeSH:
Animals;
Asia;
Asian Continental Ancestry Group*;
Biological Products;
Diagnosis;
Encephalitis Virus, Japanese*;
Encephalitis, Japanese*;
Genotype;
Humans;
Korea;
Limit of Detection;
Nervous System
- From:Journal of Bacteriology and Virology
2016;46(4):231-238
- CountryRepublic of Korea
- Language:English
-
Abstract:
Japanese encephalitis (JE) is a zoonosis that affects the nervous system of humans and other animals. The genotype of JE virus (JEV) has shifted recently from genotype 3 (G3) to genotype 1 (G1) in Asia, including Korea. Thus, a rapid differential assay is required to make an accurate diagnosis of JEV genotype. In this study, we designed common and differential primer sets for JEV G1 and G3 to detect the JEV envelope (E) gene. The specific primer sets for JEV G1 and G3 specifically amplified the target gene. The detection limits of the three primer sets were 10(1.0), 10(2.0), and 10(2.0) TCID₅₀/reaction, respectively. No cross-reactivity was detected with non-JEV reference viruses. The multiplex reverse transcription-polymerase chain reaction (RT-PCR) assay specifically differentiated JEV G1 from G3. Thus, a one-step multiplex RT-PCR assay was established to rapidly and differentially detect JEV. This assay will be useful for confirming JEV infections in animals and checking the JEV genotype in veterinary biological products.
