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- Title
Simultaneous detection of Zika, chikungunya, dengue, yellow fever, West Nile, and Japanese encephalitis viruses by a two‐tube multiplex real‐time RT‐PCR assay.
- Authors
Xu, Zhixiang; Peng, Yun; Yang, Minghui; Li, Xiaohe; Wang, Jun; Zou, Rongrong; Liang, Jinhu; Fang, Shisong; Liu, Yingxia; Yang, Yang
- Abstract
Due to the concurrent prevalence and increasing risk of coinfection of the clinically important Arboviruses, timely and accurate differential diagnosis is important for clinical management and the epidemiological investigation. A two‐tube multiplex real‐time reverse transcription‐polymerase chain reaction (RT‐PCR) assay for the simultaneous detection of Zika virus (ZIKV), chikungunya virus (CHIKV), dengue virus (DENV), yellow fever virus (YFV), West Nile virus (WNV), and Japanese encephalitis virus (JEV) was developed and optimized with high specificity and sensitivity. The detection limit for all the six viruses could reach as low as five genome equivalent copies and 2.8 × 10−3 tissue culture infectious doses (TCID50) for ZIKV, YFV, CHIKV and 2.8 × 10−2 TCID50 for JEV per reaction, with high accuracy and precision (R2 > 0.99). The coefficient of variation of intra‐assay and inter‐assay for our quantitative reverse transcription‐polymerase chain reaction (qRT‐PCR) assay was low, and the obtained positive rates ad Ct values of this assay were comparable with singleplex commercial kits. Moreover, the multiplex qRT‐PCR assay was able to detect possible co‐infections without competitive inhibition of target viral genomes. In conclusion, our rapid, sensitive, cost‐effective multiplex qRT‐PCR will be of great use for differential diagnosis in a clinical setting and epidemiological investigation during surveillance. Highlights: •A two‐tube multiplex quantitative reverse transcription‐polymerase chain reaction (qRT‐PCR) assay for the simultaneous detection of Zika virus, chikungunya virus, dengue virus, yellow fever virus, West Nile virus, and Japanese encephalitis virus was developed and optimized with high specificity and sensitivity.•The multiplex qRT‐PCR assay was able to detect possible coinfections without competitive inhibition of target viral genomes.•This assay will be of great use for the differential diagnosis in clinical setting and epidemiological investigation during surveillance.
- Subjects
JAPANESE encephalitis viruses; YELLOW fever; WEST Nile virus; CHIKUNGUNYA; DENGUE viruses
- Publication
Journal of Medical Virology, 2022, Vol 94, Issue 6, p2528
- ISSN
0146-6615
- Publication type
Article
- DOI
10.1002/jmv.27658