Sensitive and rapid detection of Zika virus by loop-mediated isothermal amplification
Zika virus (ZIKV) is a mosquito-borne flavivirus, which is a pathogen affecting humans in Africa, Asia, and America. It is necessary to detect ZIKV with a rapid and sensitive molecular method to guide timely treatment. In this study, a loop-mediated isothermal amplification (LAMP) assay was described, which is an attractive option as a fast, sensitive, and specific method for ZIKV detection using the NS5 protein coding region and the envelope protein (EP) coding region as target sequences. Two different techniques, a calcein/Mn2+ complex chromogenic method and real-time turbidity monitoring, were employed. The specificity and sensitivity of the LAMP assay were determined. The assay’s detection limit was 0.5 × 10−9 pmol/µl DNA for NS5 protein coding region and 1.12 × 10−11 pmol/µl DNA for E coding region, respectively, which is a 100-fold increase in sensitivity compared with real-time reverse transcription-polymerase chain reaction (RT-PCR) and conventional PCR. All 12 non-ZIKA respiratory pathogens tested were negative for LAMP detection, indicating the high specificity of the primers for ZIKV. In conclusion, a visual detection LAMP assay was developed, which could be a useful tool for primary quarantine purposes and clinical screening, especially in situations where resources are poor and in point-of-care tests.
KeywordsZika virus LAMP Sensitivity Specificity Rapid detection
This work received support from National Natural Science Foundation of China Grant (81401995).
JZ and RF performed the research, analyzed the data, and wrote the paper.
Compliance with ethical standards
Conflict of interest
All authors declare that they have no conflicts of interest.
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