Three pairs of primer were designed for amplification of porcine pseudorabies virus （PRV） gB, gE, and TK gene by multiplex PCR （multi-PCR） in order to differentiate vaccine strains from field isolates. Three speci...Three pairs of primer were designed for amplification of porcine pseudorabies virus （PRV） gB, gE, and TK gene by multiplex PCR （multi-PCR） in order to differentiate vaccine strains from field isolates. Three specific bands were obtained respectively at the expected size, 427 bp （gB gene）, 298 bp （gEgene）, and 208 bp （TKgene）, Then four different gene-deleted vaccines of PRV were detected by multi-PCR. One ex- pected specific band was observed in one of samples, while two bands in the others. As shown by the detection results, the multi-PCR has high sensitivity and specificity and should be applied in pathogen diagnosis and epidemiological investigation in the future.展开更多
基金Supported by National Key Technology R&D Program (2006BAD06A12)Gansu Agricultural Biotechnology Research and Application Development Project (GNSW-2005-17)~~
文摘Three pairs of primer were designed for amplification of porcine pseudorabies virus （PRV） gB, gE, and TK gene by multiplex PCR （multi-PCR） in order to differentiate vaccine strains from field isolates. Three specific bands were obtained respectively at the expected size, 427 bp （gB gene）, 298 bp （gEgene）, and 208 bp （TKgene）, Then four different gene-deleted vaccines of PRV were detected by multi-PCR. One ex- pected specific band was observed in one of samples, while two bands in the others. As shown by the detection results, the multi-PCR has high sensitivity and specificity and should be applied in pathogen diagnosis and epidemiological investigation in the future.
文摘目的建立鉴别鱼腥草Houttuynia cordata与百部还魂Gymnotheca chinensis的特异性PCR方法。方法采集不同产地的鱼腥草与百部还魂样品各8份,所有样品提取总DNA。通过对其mat K片段进行扩增、测序,进行同源比对后根据其变异位点设计特异鉴别引物,建立特异PCR鉴别方法,并通过加入SYBR Green I染料法对2种中药进行快速检测。另外,构建多重PCR体系,只经一个PCR反应,就能对鱼腥草与百部还魂进行快速分子鉴定。结果所构建特异PCR与多重PCR体系均能产生鱼腥草185 bp的特异鉴别条带,百部还魂389 bp的特异鉴别条带,SYBR Green I染料可进行快速检测方法。结论建立的鱼腥草与百部还魂2种中药的快速PCR鉴别方法简便、可靠。
