通过对荧光定量PCR反应条件的优化,建立了一种检测PCV2的SYBR Green荧光定量PCR方法。试验结果表明,该方法特异性强,与猪1型圆环病毒、猪流感病毒、猪繁殖与呼吸综合征病毒、猪伪狂犬病毒等均无交叉反应。该方法最低可检测到10拷贝/μL的DNA,比PCR检测方法敏感性高1 000倍;其标准曲线线性范围是6.84×102~6.84×107 拷贝,且具有良好的重复性。
郭慧娟
,
李秀丽
,
张国伟
,
鄢明华
,
王利丽
,
张莉
,
孙英峰
. 猪2型圆环病毒SYBR Green荧光定量PCR检测方法的建立[J]. 华北农学报, 2014
, 29(3)
: 68
-73
.
DOI: 10.7668/hbnxb.2014.03.014
By quantitative PCR optimization of reaction conditions, a real-time detection of PCV2 SYBR Green quantitative PCR method was established.The results indicated that the method was specificity, the porcine circovirus type 1(PCV1), swine influenza virus(SIV), porcine reproductive and respiratory syndrome virus(PRRSV), pig pseudorabies virus(PRV) and other common diseases of the original pig detection results were negative.The detection limit of the assay was 10 copies/μL of plasmid DNA, 1 000 times higher than that of the routine PCR.The standard curve displayed a linear range from 6.84×102 to 6.84×107 copies and a good reproducibility.
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