摘要
为探讨逆转录过程对猪繁殖与呼吸综合征病毒(PRRSV)定量检测的影响,本研究针对PRRSV保守性序列设计引物,优化反应体系及反应条件,建立准确检测PRRSV的数字PCR方法,并选择5种不同逆转录试剂盒,分析逆转录过程对数字PCR检测结果的影响。结果表明,建立的数字PCR方法线性关系良好(R^2为0.9995及0.9999),重复性良好(变异系数1.01%);利用不同逆转录试剂盒得到的定量结果存在显著性差异,揭示逆转录过程是影响RNA病毒定量检测的关键因素,提示实验过程中不同逆转录试剂盒的选用需要考察与分析。
To explore the effect of reverse transcription process on the quantitative detection of Porcine reproductive and respiratory syndrome virus(PRRSV),we designed primers based on PRRSV conservation sequences and developed a digital PCR method for accurate detection of PRRSV.The reaction systems and conditions were also optimized in the present study.Five different commercial reverse transcription kits were used to analyze the effect of reverse transcription process on the digital PCR detection.The results showed that the developed digital PCR method had good linearity(R2 was 0.9995 and 0.9999)and repeatability(coeffi cient of variation was 1.01%).The quantitative results obtained with different reverse transcription kits presented signifi cant differences,suggesting that the reverse transcription process was a key factor affecting the quantitative detection of RNA viruses.
作者
牛春艳
杨佳怡
王晶
李晶
刘文军
NIU Chun-yan;YANG Jia-yi;WANG Jing;LI Jing;LIU Wen-jun(National Institute of Metrology,China,Beijing 100029,China;Institute of Microbiology,CAS,Beijing 100101,China)
出处
《中国动物传染病学报》
CAS
北大核心
2020年第2期80-85,共6页
Chinese Journal of Animal Infectious Diseases
基金
国家重点研发计划(2016YFD0500701)。
关键词
猪繁殖与呼吸综合征病毒
RNA病毒
逆转录
数字PCR
Porcine reproductive and respiratory syndrome virus
RNA virus
reverse transcription
digital PCR
