摘要
猪圆环病毒2型(porcine circovirus type 2,PCV2)给养猪业造成了重大的经济损失,建立经济、快速的检测方法尤为重要。本研究旨在利用巢式PCR技术,建立高灵敏度PCV2检测平台,以加强对猪圆环病毒病的防控。根据PCV2 ORF2基因保守区,设计内、外2对巢式PCR引物,通过优化PCR反应条件,建立巢式PCR检测方法,并对猪场临床样本进行检测。结果显示,本研究建立的巢式PCR检测方法灵敏度高、重复性强,最低检出量为10拷贝/μL,检出率达97.3%。本研究建立的巢式PCR检测方法为快速、高效地检测PCV2以及为猪圆环病毒病的防控提供有力手段。
Porcine circovirus type 2（PCV2）caused significant economic losses of swine industry,it＇s very important to establish a economy and rapid detection method.To strengthen the prevention and control of porcine circovirus disease,rapid and high sesitive detection method of PCV2 was established using nested PCR.According to PCV2 ORF2 gene conservative region,we designed two pairs of nested PCR primers,constructed ORF2 gene recombinant plasmid for standard template and optimizated the PCR reaction conditions to establish nested PCR detection method of PCV2 and detect the clinical samples of pig farms.Our results showed that the sensitivity and repeatability of this method were very high,the minimum detection level was 10copies/μL and detection rate of clinical samples was 97.3%.In brief,the established nested PCR detection method of PCV2 in this study provided powerful means for quick and efficient detection of PCV2,and prevention and control of porcine circovirus disease in the swine industry.
出处
《中国畜牧兽医》
CAS
北大核心
2015年第12期3173-3178,共6页
China Animal Husbandry & Veterinary Medicine
基金
浙江省科技厅公益项目(2014C32061)
浙江农林大学人才启动项目(2011FR025
2013FR077)
浙江农林大学校级大学生科技创新训练计划项目(201301017)
浙江省自然科学基金项目(LQ14C010007)
关键词
猪
猪圆环病毒2型
巢式PCR
porcine
porcine circovirus type 2（PCV2）
nested PCR
