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《Chinese Journal of Blood Transfusion》 2014-06
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Comparing and application of 2 real-time PCR systems for detection herpes simplex virus in plasma donors

BI Yuedong;HU Gaoxiang;DING Xianping;College of Life Science,Sichuan University;Chengdu Institute of Biological Products. Ltd.;  
Objective To establish and evaluate the validity of 2 quantitative real-time PCR( qPCR) methods based on SYBR Green and Taqman probe for detecting herpes simplex virus( HSV). Methods According to the conserved region of UL30 gene in HSV genome obtained from GenBank,primers and Taqman probe for qPCR were designed. Recombinant plasmid which contained the segment of UL30 gene for detection was constructed as the standard template for validity,sensitivity and specificity evaluation. Also,plasma from blood products manufacturer were detected by using the established qPCR assays. Results The specificity of the established qPCR system was high and there were no cross reactivity with HBV and syphilis. The detection limit for both SYBR Green and Taqman probe qPCR system was 50 copies per reaction volume. A single melting curve peek indicated a single predominant product in SYBR Green qPCR assay. Meanwhile,in the duplicated experiment,the replication of intra-assay and inter-assay was good. Additionally,the qPCR systems were further successfully applied to detect the viral load of HSV DNA in plasma from donors with the recombinant vector as the standard temples. Two methods were used to detecte HSV in plasma domor,which detected 2. 08%( 3 /144) of the specimen were positive for HSV DNA. Conclusion With the characteristics of high sensitivity,strong specificity,and good repeatability,the 2 qPCR systems can be used for HSV infection detection in blood samples in order to improve the blood safety in China.
【CateGory Index】: R373.11;R3416
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Chinese Journal Full-text Database 10 Hits
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