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Development of one step real-time RT-PCR method for detection of West Nile virus

Peng Xie

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Abstract

One step real-time RT-PCR(Taqman) assays of West Nile virus(WNV) envelope protein gene was developed so that WNV RNA in field specimens or laboratory material can be characterized rapidly and specifically.A pair of specific oligonucleotide primers was designed.The linearity of the standard curve allowed quantification of 102 to 107 RNA transcripts/μl.and sensitivity of the test was close to 3.6×102transcripts/μl.This assays were high specific without cross-reaction to Japanese encephalitis virus or other virus.This standardized technique is sensitive and reliable and allows rapid detection and quantitation of West Nile virus RNA in both field and experimental materials used for the surveillance and specific diagnosis of West Nile virus.

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What this paper is about

One step real-time RT-PCR(Taqman) assays of West Nile virus(WNV) envelope protein gene was developed so that WNV RNA in field specimens or laboratory material can be characterized rapidly and specifically.A pair of specific oligonucleotide primers was designed.The linearity of the standard curve allowed quantification of 102 to 107 RNA transcripts/μl.and sensitivity of the test was close to 3.6×102transcripts/μl.This assays were high specific without cross-reaction to Japanese encephalitis virus or other virus.This standardized technique is sensitive and reliable and allows rapid detection and quantitation of West Nile virus RNA in both field and experimental materials used for the surveillance and specific diagnosis of West Nile virus.

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Available abstract

One step real-time RT-PCR(Taqman) assays of West Nile virus(WNV) envelope protein gene was developed so that WNV RNA in field specimens or laboratory material can be characterized rapidly and specifically.A pair of specific oligonucleotide primers was designed.The linearity of the standard curve allowed quantification of 102 to 107 RNA transcripts/μl.and sensitivity of the test was close to 3.6×102transcripts/μl.This assays were high specific without cross-reaction to Japanese encephalitis virus or other virus.This standardized technique is sensitive and reliable and allows rapid detection and quantitation of West Nile virus RNA in both field and experimental materials used for the surveillance and specific diagnosis of West Nile virus.

Key concepts: Virology, West Nile virus, TaqMan, Biology, Virus, Flavivirus, Real-time polymerase chain reaction, RNA

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