【摘要】 为建立快速检测猪圆环病毒2型(PCV2)的LAMP方法,根据GenBank发表的PCV2ORF2基因组保守序列,设计合成4条特异性LAMP引物,通过PCV2ORF2的制备、反应条件的优化、敏感性试验和特异性试验,初步建立了PCV2LAMP检测方法,并对60份临床检样和6株PCV2进行了检测应用。结果显示,建立的PCV2LAMP检测方法具有良好的特异性和敏感性,在64℃45min条件下可扩增出大于134bp的特异性梯状DNA条带,检测限度可达10copies/μL;用它对PCV1、PRV、PPV、CSFV和PRRSV的核酸进行扩增,但结果均为阴性。用建立的LAMP方法进行的样品检测结果显示,6株受检PCV2毒株的检出率为100%,60份临床检样PCV2的阳性率为20%(12/60),与PCR方法平行检测的结果相一致。更多还原

【Abstract】 To develop a loop-mediated isothermal amplification(LAMP)method for quick detection of porcine circovirus type 2(PCV2),4specific LAMP primers were designed and synthesized according to the conserved sequences of ORF2gene of PCV2published in GenBank.The LAMP assay was established and evaluated by the preparation of target gene fragments,optimization of reaction conditions,sensitivity and specificity tests.Then 6strains of PCV2and 60clinical samples were detected using LAMP and PCR in parallel.In result,the LAMP method for the PCV2detection performed at 64℃for 45min showed a ladder-like pattern of amplification bands more than 134bp.The detection limitation reached 10copies/μL,and the assay gave no amplification from PCV1,PRV,PPV,CSFV and PRRSV.PCV2positive rate in the detected samples was 20%and all the 6strains of PCV2were positive by the two methods.These results indicated the good correlation between PCV2LAMP and PCR.更多还原