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Development of multiplex real‐time reverse‐transcriptase polymerase chain reaction assay for simultaneous detection of Zika, dengue, yellow fever, and chikungunya viruses in a single tube
Author(s) -
Wu Wenhui,
Wang Jin,
Yu Nan,
Yan Juying,
Zhuo Zhihao,
Chen Mengyuan,
Su Xiaosong,
Fang Mujin,
He Shuizhen,
Zhang Shiyin,
Zhang Yanjun,
Ge Shengxiang,
Xia Ningshao
Publication year - 2018
Publication title -
journal of medical virology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.782
H-Index - 121
eISSN - 1096-9071
pISSN - 0146-6615
DOI - 10.1002/jmv.25253
Subject(s) - virology , chikungunya , zika virus , dengue virus , dengue fever , virus , biology , multiplex , yellow fever , reverse transcriptase , polymerase chain reaction , gene , bioinformatics , biochemistry
Zika virus (ZIKV), dengue virus (DENV), chikungunya virus (CHIKV) and yellow fever virus (YFV) share the same mosquito vectors and have similar clinical manifestations early stage of infection. Therefore, simultaneously differentiating these viruses from each other is necessary. We developed a multiplex real‐time reverse‐transcriptase polymerase chain reaction (RT‐PCR) assay for the differentiation of these four viruses in a single tube. The linear range was established by regression analysis, and the R 2 value for each virus was ≥0.98, and the 95% lower limit of detection for each virus was as follows (copies/reaction): ZIKV‐Asian, 9; ZIKV‐Africa, 15; CHIKV, 11; DENV‐1, 19; DENV‐2, 13; DENV‐3, 24; DENV‐4, 36; and YFV, 17. Meanwhile, our multiplex real‐time RT‐PCR has a good consistency with the commercial singleplex assay. In summary, the developed assay can be effectively used for the diagnosis of ZIKV, DENV, CHIKV, and YFV infections.