Comparative Study on Three Methods of Molecular Biology for Detecting Orf Virus
Design LAMP primers(F3&B3,BIP & FIP)for the gene Orf Virus(OrfV)B2L,establish detection methods of LAMP,use outer primers(F3&B3)of LAMP,set up two conventional molecular detection methods-PCR and RTFQ-PCR respectively,compare and study the specificity,sensitivity,detection coincidence rate and time limit of the three detection methods.The results showed that the three methods only conduct special amplification to nucleic acid of OrfV but not to Goat pox virus(GTPV),foot-and-mouth disease virus(FMDV),Peste des Petits Ruminants virus(PPRV)and extraction of nucleic acids from the skin of healthy goats; LAMP method for detecting of OrfV nucleic acid template has the lowest detection limit of 1.0×10-8 dilution(5.3 fg),detection time limit of about 90 min,and the method RTFQ-PCR has the lowest detection limit of 1.0×104 copies/μL(18.2 fg),detection time limit of about 120 min while the conventional method PCR has the lowest detection limit of 1.0×10-5 dilution(5.3 pg)and the detection time limit of about 150 min.After conversion,the sensitivity of LAMP is equivalent to that of RTFQ-PCR and their sensitivities are 1000 times higher than that of conventional PCR.5 clinical cases of OrfV infection were detected by three methods with the positive rate of 100%and the coincidence rate of 100%.Comparative study on three methods for detecting
Orf Virus (OrfV) LAMP Real-time fluorescent quantitative PCR conventional PCR
XIAN Si-mei LI Peng-fei FENG Jiang LIU Ai YANG Yu LIU Zong-sheng
College of Animal Science,Guizhou University,Gui yang 550025,Gui zhou,china;key Laboratory of Animal College of Animal Science,Guizhou University,Gui yang 550025,Gui zhou,china Institute of Supervision and Inspection for Animal Products Quality and Safety,Bijie 551700,china
国内会议
重庆
英文
1285-1296
2016-05-25（万方平台首次上网日期，不代表论文的发表时间）