Comparative Study on Three Methods of Molecular Biology for Detecting Orf Virus
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- 英文论文题名：Comparative Study on Three Methods of Molecular Biology for Detecting Orf Virus
- 论文作者：Xian Simei ; Li Pengfei ; Feng Jiang ; Liu Ai ; Yang Yu ; Liu Zongsheng
- 英文论文作者：Xian Simei ; Li Pengfei ; Feng Jiang ; Liu Ai ; Yang Yu ; Liu Zongsheng ; College of Animal Science ; Guizhou University ; Key Laboratory of Animal Disease and Veterinary Public Health of Gui zhou Province ; Institute of Supervision and Inspection for Animal Products Quality and Safety
- 年：2016
- 作者机构：College of Animal Science,Guizhou University;Key Laboratory of Animal Disease and Veterinary Public Health of Gui zhou Province;Institute of Supervision and Inspection for Animal Products Quality and Safety,Bijie;
- 英文论文关键词：Orf Virus(OrfV) ; LAMP ; Real-time fluorescent quantitative PCR ; conventional PCR
- 会议召开时间：2016-05-25
- 会议录名称：第七届中国畜牧科技论坛论文集
- 英文会议录名称：Proceeding of the 7th China Animal Husbandry Sci-tech Forum
- 语种：英文
- 分类号：S852.65
- 学会代码：ZGXJ
- 会议名称：第七届中国畜牧科技论坛
- 会议地点：中国重庆
- 主办单位：中国畜牧兽医学会
- 学会名称：中国畜牧兽医学会
- 页数：1
- 文件大小：48k
- 原文格式：O
- 会议级别：全国
摘要
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 1.0×10~4copies/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 are1000 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 OrfV with the same pair of primers was established,and the LAMP method becomes the molecular detection means that is suitable for basic level application and promotion with high sensitivity,strong specificity,high-speed and highefficiency,low equipment requirements.
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 1.0×10~4copies/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 are1000 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 OrfV with the same pair of primers was established,and the LAMP method becomes the molecular detection means that is suitable for basic level application and promotion with high sensitivity,strong specificity,high-speed and highefficiency,low equipment requirements.
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 1.0×10~4copies/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 are1000 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 OrfV with the same pair of primers was established,and the LAMP method becomes the molecular detection means that is suitable for basic level application and promotion with high sensitivity,strong specificity,high-speed and highefficiency,low equipment requirements.
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