猪圆环病毒3型PCR诊断方法的建立
发布时间:2018-04-30 22:39
本文选题:猪圆环病毒型 + 聚合酶链式反应 ; 参考:《中国兽医科学》2017年12期
【摘要】:根据NCBI中PCV3的基因组序列,设计了1对特异性引物,并对PCR的反应体系和反应条件进行优化,同时进行特异性和敏感性试验,最后应用所建立的PCR方法检测了临床送检的22份病料。结果显示,建立的PCR方法的最佳反应条件为总体积25 L,退火温度56℃,模板DNA量3 L(l ng/m L),进行35个循环。该PCR能特异地扩增出目的片段,长度约为381 bp。测序结果表明,PCR扩增产物正确。该PCR反应特异性强,只能扩增PCV3阳性样品,且能扩增的最低模板量为1×10-4ng/m L。用该PCR对送检的22份病料进行PCV3检测,结果阳性率为13.6%。可见,本研究建立的PCR方法特异、敏感,可用于PCV3的快速检测。
[Abstract]:According to the genomic sequence of PCV3 in NCBI, a pair of specific primers was designed, and the reaction system and reaction conditions of PCR were optimized, and the specificity and sensitivity tests were carried out. Finally, 22 samples were detected by the established PCR method. The results show that the optimum reaction conditions of the PCR method are: total volume 25 L, annealing temperature 56 鈩,
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