tetano
Editor, Senior Moderator
Zhonghua Shi Yan He Lin Chuang Bing Du Xue Za Zhi. 2010 Oct;24(5):383-5.
[The study of multiple RT-PCR-based reverse dot blot hybridization technique for detecting influenza viruses].
[Article in Chinese]
Yang L, Zhang XM, Zhang XG, Ma J, Wang M, Wen LY, Wang DY, Bai T, Shu YL, Qian YH, Zeng Y.
Abstract
OBJECTIVE: To establish a multiplex RT-PCR-based reverse dot blot hybridization technique to detect influenza viruses.
METHODS: Obtain the HA nucleotide sequences of seasonal influenza H1N1, seasonal influenza H3N2, influenza H1N1 and human avian influenza H5N1 from GenBank. Design primers in conservative district and probes t in high variable region respectively, after analyzing the HA nucleotide sequences of influenza virus through the Vector NTI 9.0. Establish and optimize multiple RT-PCR system by comparing amplification efficiency and specificity at different primer concentrations. Establish the reverse dot hybridization system after optimizing the concentration of probes. To compare the sensitivity and specificity of this technique and the general RT-PCR Method through extracting the viral RNA of the mentioned influenza virus which are to be the reference substance.
RESULTS: Successfully establish a multiplex RT-PCR-based reverse dot blot hybridization technique for detecting influenza viruses. This technique is 100-1000 times more sensitive than gel electrophoresis method, and it has a good specificity.
CONCLUSION: Successfully established multiplex RT-PCR-based reverse dot blot hybridization technique for detecting influenza viruses.
PMID: 21280332 [PubMed - in process]
http://www.ncbi.nlm.nih.gov/pubmed/21280332
[The study of multiple RT-PCR-based reverse dot blot hybridization technique for detecting influenza viruses].
[Article in Chinese]
Yang L, Zhang XM, Zhang XG, Ma J, Wang M, Wen LY, Wang DY, Bai T, Shu YL, Qian YH, Zeng Y.
Abstract
OBJECTIVE: To establish a multiplex RT-PCR-based reverse dot blot hybridization technique to detect influenza viruses.
METHODS: Obtain the HA nucleotide sequences of seasonal influenza H1N1, seasonal influenza H3N2, influenza H1N1 and human avian influenza H5N1 from GenBank. Design primers in conservative district and probes t in high variable region respectively, after analyzing the HA nucleotide sequences of influenza virus through the Vector NTI 9.0. Establish and optimize multiple RT-PCR system by comparing amplification efficiency and specificity at different primer concentrations. Establish the reverse dot hybridization system after optimizing the concentration of probes. To compare the sensitivity and specificity of this technique and the general RT-PCR Method through extracting the viral RNA of the mentioned influenza virus which are to be the reference substance.
RESULTS: Successfully establish a multiplex RT-PCR-based reverse dot blot hybridization technique for detecting influenza viruses. This technique is 100-1000 times more sensitive than gel electrophoresis method, and it has a good specificity.
CONCLUSION: Successfully established multiplex RT-PCR-based reverse dot blot hybridization technique for detecting influenza viruses.
PMID: 21280332 [PubMed - in process]
http://www.ncbi.nlm.nih.gov/pubmed/21280332