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Development and application of a real-time fluorescence quantitative PCR method for detecting West Nile virus

Shi Yu-lin

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Abstract

Objective To develop a real-time fluorescence quantitative PCR method for detecting West Nile virus. Methods Primers were designed according to the conserved sequence of the envelope protein (E) of West Nile virus. SYBR Green-based real-time PCR was established. The specificity, sensitivity and stability of the method were assessed. 54 clinical specimens were detected by the established SYBR Green-based real-time PCR. Results High specificity was found using this established method to detect West Nile virus and no cross reactions were observed for other viruses such as JEV and SLEV. The sensitivity of this method was low to 100 pg. The correlation coefficient and slope value of standard curve were 0.995 and -3.444, respectively. The accuracy of clinical diagnosis was 98.15% for 54 clinical specimens. Conclusion Real time quantitative PCR was a reliable, specific, sensitive and useful tool for detecting West Nile virus.

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What this paper is about

Objective To develop a real-time fluorescence quantitative PCR method for detecting West Nile virus. Methods Primers were designed according to the conserved sequence of the envelope protein (E) of West Nile virus. SYBR Green-based real-time PCR was established. The specificity, sensitivity and stability of the method were assessed. 54 clinical specimens were detected by the established SYBR Green-based real-time PCR. Results High specificity was found using this established method to detect West Nile virus and no cross reactions were observed for other viruses such as JEV and SLEV. The sensitivity of this method was low to 100 pg. The correlation coefficient and slope value of standard curve were 0.995 and -3.444, respectively. The accuracy of clinical diagnosis was 98.15% for 54 clinical specimens. Conclusion Real time quantitative PCR was a reliable, specific, sensitive and useful tool for detecting West Nile virus.

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Available abstract

Objective To develop a real-time fluorescence quantitative PCR method for detecting West Nile virus. Methods Primers were designed according to the conserved sequence of the envelope protein (E) of West Nile virus. SYBR Green-based real-time PCR was established. The specificity, sensitivity and stability of the method were assessed. 54 clinical specimens were detected by the established SYBR Green-based real-time PCR. Results High specificity was found using this established method to detect West Nile virus and no cross reactions were observed for other viruses such as JEV and SLEV. The sensitivity of this method was low to 100 pg. The correlation coefficient and slope value of standard curve were 0.995 and -3.444, respectively. The accuracy of clinical diagnosis was 98.15% for 54 clinical specimens. Conclusion Real time quantitative PCR was a reliable, specific, sensitive and useful tool for detecting West Nile virus.

Key concepts: West Nile virus, Real-time polymerase chain reaction, Virology, Melting curve analysis, Virus, Biology, SYBR Green I, Nile red

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