Micro-droplet Digital Polymerase Chain Reaction and Real-Time Quantitative Polymerase Chain Reaction Technologies Provide Highly Sensitive and Accurate Detection of Zika Virus

作者：Hui, Yuan; Wu, Zhiming; Qin, Zhiran; Zhu, Li; Liang, Junhe; Li, Xujuan; Fu, Hanmin; Feng, Shiyu; Yu, Jianhai; He, Xiaoen; Lu, Weizhi; Xiao, Weiwei; Wu, Qinghua; Zhang, Bao^*; Zhao, Wei^*

来源：VIROLOGICA SINICA, 2018, 33(3): 270-277.

DOI：10.1007/s12250-018-0037-y

• Zika virus
• Nucleic acid detection
• Micro-droplet digital polymerase chain reaction (ddPCR)
• Real-time fluorescence quantitative polymerase chain reaction (RT-qPCR)

The establishment of highly sensitive diagnostic methods is critical in the early diagnosis and control of Zika virus (ZIKV) and in preventing serious neurological complications of ZIKV infection. In this study, we established micro-droplet digital polymerase chain reaction (ddPCR) and real-time quantitative PCR (RT-qPCR) protocols for the detection of ZIKV based on the amplification of the NS5 gene. For the ZIKV standard plasmid, the RT-qPCR results showed that the cycle threshold (Ct) value was linear from 10(1) to 10(8) copy/mu L, with a standard curve R-2 of 0.999 and amplification efficiency of 92.203%; however, a concentration as low as 1 copy/mu L could not be detected. In comparison with RT-qPCR, the ddPCR method resulted in a linear range of 10(1)-10(4) copy/mu L and was able to detect concentrations as low as 1 copy/mu L. Thus, for detecting ZIKV from clinical samples, RT-qPCR is a better choice for high-concentration samples (above 10(1) copy/mu L), while ddPCR has excellent accuracy and sensitivity for low-concentration samples. These results indicate that the ddPCR method should be of considerable use in the early diagnosis, laboratory study, and monitoring of ZIKV.

• 出版日期2018-6
• 单位南方医科大学