摘要
根据PCV2 ORF1设计1对特异性引物,建立了SYBR GreenⅠReal-time PCR检测方法。该方法灵敏度可达10-100拷贝/μL,比常规PCR检测方法灵敏度高10倍,而与猪繁殖和呼吸综合征病毒（PRRSV）、猪伪狂犬病毒（PRV）、猪瘟病毒（CSFV）以及猪细小病毒（PPV）没有交叉反应,利用该方法从50份临床病料中检测出43份阳性PCV2病毒,阳性检出率为94%。与常规PCR比较结果显示,该方法具有较高的灵敏度和特异性,能够快速检测和监控PCV2的感染流行。
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 synthesized 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 pseudorabies virus（ PRV）. The limits of detection and quantitation were 10 and 100 copies,respectively. Using the established 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.
出处
《华北农学报》
CSCD
北大核心
2014年第2期66-70,共5页
Acta Agriculturae Boreali-Sinica
基金
中国博士后科学基金项目(2011M501179)
河南省教育厅科学技术研究重点项目(13A230838)
