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

Wenhui Wu; Jin Wang; Nan Yu; Juying Yan; Zhihao Zhuo; Mengyuan Chen; Xiaosong Su; Mujin Fang; Shuizhen He; Shiyin Zhang; Yanjun Zhang; Shengxiang Ge; Ningshao Xia

doi:10.1002/jmv.25253

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

J Med Virol. 2018 Nov;90(11):1681-1686. doi: 10.1002/jmv.25253. Epub 2018 Aug 16.

Authors

Wenhui Wu^{1

2}, Jin Wang^{1

2}, Nan Yu³, Juying Yan⁴, Zhihao Zhuo^{1

2}, Mengyuan Chen^{1

2}, Xiaosong Su^{1

2}, Mujin Fang^{1

2}, Shuizhen He⁵, Shiyin Zhang^{1

2}, Yanjun Zhang⁴, Shengxiang Ge^{1

2}, Ningshao Xia^{1

2}

Affiliations

¹ National Institute of Diagnostics and Vaccine Development in Infectious Diseases, School of Life Sciences, Xiamen University, Xiamen, China.
² State Key Laboratory of Molecular Vaccinology and Molecular Diagnostics, School of Public Health, Xiamen University, Xiamen, China.
³ Center of Clinical Laboratory Diagonosis, Zhujiang Hospital, Southern Medical University, Guangzhou, China.
⁴ Zhejiang Center for Disease Control and Prevention, Hangzhou, China.
⁵ Xiamen Center for Disease Control and Prevention, Xiamen, China.

PMID: 29979812
DOI: 10.1002/jmv.25253

Abstract

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 R² 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.

Keywords: Zika; chikungunya; dengue; multiplex real-time reverse-transcriptase polymerase chain reaction; yellow fever.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Africa
Chikungunya Fever / diagnosis*
Chikungunya Fever / virology
Chikungunya virus / genetics
Chikungunya virus / isolation & purification
Dengue / diagnosis*
Dengue / virology
Dengue Virus / genetics
Dengue Virus / isolation & purification
Humans
Molecular Diagnostic Techniques / methods
Multiplex Polymerase Chain Reaction / methods*
Real-Time Polymerase Chain Reaction / methods*
Reverse Transcriptase Polymerase Chain Reaction / methods*
Time Factors
Yellow Fever / diagnosis*
Yellow Fever / virology
Yellow fever virus / genetics
Yellow fever virus / isolation & purification
Zika Virus / genetics
Zika Virus / isolation & purification
Zika Virus Infection / diagnosis*
Zika Virus Infection / virology

Abstract

Publication types

MeSH terms

Grants and funding