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Wei Sheng Yan Jiu. 2012 Jul;41(4):662-5.
[Rapid detection of the N1 and N2 subtype of influenza virus by duplex fluorescence quantitative RT-PCR].
[Article in Chinese]
Huang L, Zhang H, Wang T, Fang S, Wang X, Li J, Cheng X, Lv X, Wu C, Zhang R, Cheng J.
Abstract
OBJECTIVE:
To develop a method to rapidly detect the N1 and N2 subtype of influenza virus by fluorescence real-time quantitative PCR.
METHODS:
According to the conservative sequences of NA gene of the N1 and N2 subtype of influenza virus, two pairs of primer and Taq-man probe were designed, respectively. Using one step RT-PCR kit to set up a duplex RT-PCR system for detecting the N1 and N2 subtype of influenza virus; the standard quantitative curve of the assay was established by using 10-fold serial dilution of in-vitro transcribed RNA, and the sensitivity and reproducibility was determined. The specificity and test for influenza virus and respirovirus were also determined by using this duplex Real-time RT-PCR system.
RESULTS:
The sensitivity of detecting the N1 and N2 subtype of influenza virus was 10 copies/microl, and the regression coefficient of the quantitative curve was higher than 99%. The amplification efficiency and specificity of this assay was 102.21% and 101.78% respectively. The system was capable of detecting human influenza viruses with high specificity.
CONCLUSION:
The detection system based on RT-PCR could be utilized to rapidly and sensitively detect the N1 and N2 subtype of influenza virus.
PMID:
23057336
[PubMed - in process]
[Rapid detection of the N1 and N2 subtype of influenza virus by duplex fluorescence quantitative RT-PCR].
[Article in Chinese]
Huang L, Zhang H, Wang T, Fang S, Wang X, Li J, Cheng X, Lv X, Wu C, Zhang R, Cheng J.
Abstract
OBJECTIVE:
To develop a method to rapidly detect the N1 and N2 subtype of influenza virus by fluorescence real-time quantitative PCR.
METHODS:
According to the conservative sequences of NA gene of the N1 and N2 subtype of influenza virus, two pairs of primer and Taq-man probe were designed, respectively. Using one step RT-PCR kit to set up a duplex RT-PCR system for detecting the N1 and N2 subtype of influenza virus; the standard quantitative curve of the assay was established by using 10-fold serial dilution of in-vitro transcribed RNA, and the sensitivity and reproducibility was determined. The specificity and test for influenza virus and respirovirus were also determined by using this duplex Real-time RT-PCR system.
RESULTS:
The sensitivity of detecting the N1 and N2 subtype of influenza virus was 10 copies/microl, and the regression coefficient of the quantitative curve was higher than 99%. The amplification efficiency and specificity of this assay was 102.21% and 101.78% respectively. The system was capable of detecting human influenza viruses with high specificity.
CONCLUSION:
The detection system based on RT-PCR could be utilized to rapidly and sensitively detect the N1 and N2 subtype of influenza virus.
PMID:
23057336
[PubMed - in process]
