华北农学报Issue(2)：66-70,5.
猪圆环病毒2型荧光定量PCR检测方法的建立
Rapid Detection of Porcine Circovirus Type 2 Using a SYBR Green Ⅰ Real-time PCR
摘要
Abstract
Porcine circovirus type 2 ( PCV2 ) is the causative factor of Post-weaning multi-systemic wasting syndrome ( PMWS) in pigs. Rapid detection of PCV2 is very important for the effective prophylaxis and treatment of PMWS. To establish a sensitive,specific assay for the detection and quantitation of PCV2,we designed and synthe-sized specific primers in the open reading frame 1(ORF1),and to develop a SYBR Green ⅠReal-time PCR. The results indicated that the Real-time PCR assay could detect 10-100 copies of the genomic DNA per reaction,and its sensitivity was 10 times of the conventional PCR. The assay did not cross-react with classical swine fever virus ( CSFV) ,porcine parvovirus( PPV) ,porcine reproductive and respiratory syndrome virus ( PRRSV) and pseudora-bies virus (PRV). The limits of detection and quantitation were 10 and 100 copies,respectively. Using the estab-lished Real-time PCR system,43 of the 50 samples we tested were detected as positive. In conclusion,the Real-time PCR assay is sensitive,specific,accurate and can be used for the monitoring of PCV2 infection.关键词
猪圆环病毒2型/SYBR Green Ⅰ/荧光定量PCRKey words
Porcine circovirus type 2/SYBR Green Ⅰ/Real-time PCR分类
生物科学引用本文复制引用
李鹏,郭军庆,金前跃,李青梅,李清州,万博,王选年,王川庆,张改平..猪圆环病毒2型荧光定量PCR检测方法的建立[J].华北农学报,2014,(2):66-70,5.基金项目
中国博士后科学基金项目(2011M501179) （2011M501179）
河南省教育厅科学技术研究重点项目(13A230838) （13A230838）
