The detection of 10 violent RNA viruses by microarray technology
- VernacularTitle:基因芯片技术检测10种烈性RNA病毒
- Author:
Yinhui YANG
;
Ruifu YANG
;
Guohui CHANG
- Publication Type:Journal Article
- Keywords:
oligonucleotide array sequence analysis;
RNA, viral;
detection
- From:
Medical Journal of Chinese People's Liberation Army
1982;0(03):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective To detect 10 RNA viruses including Alphavirus in Togoviridae, Flavivirus in Flaviridae, Hantavirus and Nairovirus in Bunyavirudae and SARS-CoV in Coronavirudae by using genechip technique. Methods The universal PCR primers of Alphavirus and Flavivirus and the PCR primers specific for HFRSV in Hantavirus and XJHFV in Nairovirus were designed by DNAStar software. PCR primers specific for SARS-CoV were adopted from WHO website. In addition, all the PCR primers specific for each virus were designed inside the regions of universal primers. These specific primers were utilized for amplification of cDNA probes. The concentration of probes, the hybridization temperature and duration, the formulation of hybridization solution and the washing conditions were optimized. Results The specific hybridization signals could be obtained when the concentration of probes was 0.3?g/?l. Good hybridization signals could be obtained for all the 10 RNA viruses when the hybridization solution contained 20% formamide, and the hybridization reaction was conducted at 60℃ for 1.5 hours. Two or four pathogens could be detected simultaneously when the target nuclear acids were amplified by multiplex PCR. Conclusion The results showed that the virus pathogens could be detected by genechip technique, and the key step was to design suitable primers and probes.
