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Detection and Differentiation of Genotype I and III Japanese Encephalitis Virus in Mosquitoes by Multiplex Reverse Transcriptase‐Polymerase Chain Reaction
Author(s) -
Chen Y. Y.,
Lin J. W.,
Fan Y. C.,
Chiou S. S.
Publication year - 2014
Publication title -
transboundary and emerging diseases
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.392
H-Index - 63
eISSN - 1865-1682
pISSN - 1865-1674
DOI - 10.1111/j.1865-1682.2012.01369.x
Subject(s) - primer (cosmetics) , virology , multiplex , japanese encephalitis , multiplex polymerase chain reaction , biology , reverse transcriptase , genotype , virus , polymerase chain reaction , microbiology and biotechnology , gene , encephalitis , genetics , chemistry , organic chemistry
Summary Japanese encephalitis ( JE ) is a disease that threatens both human and animal populations in A sian countries, and the causative agent of JE , J apanese encephalitis virus ( JEV ), has recently changed from genotype III ( GIII ) to genotype I ( GI ). However, a test for the rapid differentiation of GI and GIII JEV is still unavailable, especially one that can be used for mosquito‐based surveillance. We have designed GI ‐ and GIII ‐specific primer sets for the rapid detection and differentiation of GI and GIII JEV by multiplex reverse transcriptase‐polymerase chain reaction (multiplex RT ‐ PCR ). The GI ‐specific and GIII ‐specific primer sets were able to specifically amplify the target gene from GI and GIII JEV , respectively. The limitations of detection were 0.00225 and 0.225 pfu for the GI ‐specific and GIII ‐specific primers, respectively. Using a mixture of GI ‐specific and GIII ‐specific primers, the multiplex RT ‐ PCR was able to specifically detect and differentiate GI and GIII JEV . The multiplex RT ‐ PCR was able to successfully differentiate GI and GIII virus in JEV ‐infected mosquitoes. Thus, a sensitive and specific multiplex RT ‐ PCR system for the rapid detection and differentiation of GI and GIII JEV has been developed, and this test is likely to be valuable when carrying out mosquito‐based JEV surveillance.