Research Article: Rapid and reliable diagnostic method to detect Zika virus by real-time fluorescence reverse transcription loop-mediated isothermal amplification

Date Published: April 18, 2018

Publisher: Springer Berlin Heidelberg

Author(s): Xu-Guang Guo, Yong-Zhuo Zhou, Qin Li, Wei Wang, Jin-Zhou Wen, Lei Zheng, Qian Wang.


To detect Zika virus more rapidly and accurately, we developed a novel method that utilized a real-time fluorescence reverse transcription loop-mediated isothermal amplification (LAMP) technique. The NS5 gene was amplified by a set of six specific primers that recognized six distinct sequences. The amplification process, including 60 min of thermostatic reaction with Bst DNA polymerase following real-time fluorescence reverse transcriptase using genomic Zika virus standard strain (MR766), was conducted through fluorescent signaling. Among the six pairs of primers that we designate here, NS5 was the most efficient with a high sensitivity of up to 3.3 ng/μl and reproducible specificity on eight pathogen samples that were used as negative controls. The real-time fluorescence reverse transcription LAMP detection process can be completed within 35 min. Our study demonstrated that real-time fluorescence reverse transcription LAMP could be highly beneficial and convenient clinical application to detect Zika virus due to its high specificity and stability.

Partial Text

Zika virus is primarily transmitted by the bite of the female Aedes aegypti mosquito in tropical regions, which must feed on blood to lay eggs (Gulland 2016; Walsh 2016). Other species of mosquito have been reported such as Aedes polynesiensis and Aedes albopictus. The virus has also been isolated from a number of arboreal mosquito species in the Aedes genus.

The Zika virus belongs to the Flaviviridae family and the Flavivirus genus and is thus related to the dengue fever, yellow fever, Japanese encephalitis, and West Nile viruses (Zuanazzi et al. 2017). Most patients show no symptoms or have mild symptoms (Zhang et al. 2017). Only 20% of patients experience fever, rash, conjunctivitis and joint pain, while in some cases, symptoms disappear within a week (Wilbe et al. 2017; Zamani and Zamani 2017). Therefore, to develop a rapid and robust nucleic acid amplification assay to efficiently detect Zika virus is critically important for clinical treatment and disease prevention.




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