Establishment of methods for detection of dengue virus by real-time quantitative RT-PCR with SYBR Green I.
Bing Wu
Abstract
Bing Wu
Abstract
Objective To develop a real-time quantitative RT-PCR method with SYBR Green I to detect dengue virus. Methods The optimal conditions and system of real-time PCR using SYBR Green I were established.The sensitivity and specificity and repeat were analyzed. Results The size of amplified products of the standard virus was obtained as expected.The sequencing results revealed that the amplified products sequences consisted of the standard dengue sequences in NCBI.The sensitivity,specificity and reproductivity of SYBR Green I real-time PCR was high.The specification of amplified products was checked by melting curve analysis. Conclusion The established SYBR Green I real-time PCR is a rapid,specific and sensitive method for the detection of Dengue virus.It can be used for monitoring of early dengue infection.
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Objective To develop a real-time quantitative RT-PCR method with SYBR Green I to detect dengue virus. Methods The optimal conditions and system of real-time PCR using SYBR Green I were established.The sensitivity and specificity and repeat were analyzed. Results The size of amplified products of the standard virus was obtained as expected.The sequencing results revealed that the amplified products sequences consisted of the standard dengue sequences in NCBI.The sensitivity,specificity and reproductivity of SYBR Green I real-time PCR was high.The specification of amplified products was checked by melting curve analysis. Conclusion The established SYBR Green I real-time PCR is a rapid,specific and sensitive method for the detection of Dengue virus.It can be used for monitoring of early dengue infection.
Key concepts: SYBR Green I, Real-time polymerase chain reaction, Dengue fever, Dengue virus, Melting curve analysis, Virology, Biology, Virus