[Establishment and application of fluorescent quantitative PCR method for detecting human herpes virus type 6].
作者信息
Dong Xue-jun, Shi Chang-gen, Qu Shi-wen, Liu Zhong-min, Fu Jia-ping
机构信息
Central Laboratory, Shaoxing People's Hospital, Shaoxing, China.
出版信息
Zhonghua Shi Yan He Lin Chuang Bing Du Xue Za Zhi. 2005 Mar;19(1):74-6.
OBJECTIVE
To establish fluorescent quantitative PCR method for detecting human herpes virus type 6 (HHV6).
METHODS
According to the specific sequence of human herpes virus type 6 genes, the primers and the fluorescent probe (TaqMan) were designed and synthesized. The fragment generated from HHV 6 gene as template was cloned into the pMD18-T vector which was constructed from the pUC 18. And the positive recombinant plasmid was 1:10 diluted and used as standard quantitative template to make the standard curve for sample detection.
RESULTS
The standard curve indicated the linear relationship between Ct (cycle threshold) and template concentration. The clinical samples from 135 cases were detected by this system, 16 cases among 135 were positive.
CONCLUSION
The fluorescent quantitative PCR method for the detection of human herpes virus type 6 is simple and accurate, and this method may be helpful to clinical diagnosis.
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