綿羊痘病毒TaqMan-MGB熒光定量PCR快速檢測方法的建立
發(fā)布時間:2018-08-16 13:52
【摘要】:為建立快速檢測綿羊痘病毒(SPPV)快速檢測方法基因的方法,本研究參照SPPV ORF068基因設(shè)計并合成一對特異性引物和一條MGB探針,經(jīng)條件優(yōu)化,建立了SPPV TaqMan-MGB熒光定量PCR快速檢測方法。結(jié)果顯示,該方法可以特異性檢測SPPV,而對羊口瘡病毒和豬水皰病病毒等擴增結(jié)果均為陰性;而且檢出下限為10拷貝/μL;組內(nèi)組間重復性試驗的變異系數(shù)均小于3%。上述結(jié)果顯示本研究建立的TaqMan-MGB qPCR方法特異性強、靈敏度高、重復性好,能夠?qū)PPV進行快速準確的檢測。
[Abstract]:In order to establish a rapid detection method for the (SPPV) gene of sheep pox virus, a pair of specific primers and a MGB probe were designed and synthesized according to the SPPV ORF068 gene in this study. The rapid detection method of SPPV TaqMan-MGB fluorescence quantitative PCR was established by optimizing the conditions. The results showed that this method could be used to detect SPPV-specific, but negative for sheep aphtha virus and porcine blister virus, and the detection limit was 10 copies / 渭 L, and the coefficient of variation of repeated test was less than 3. The above results show that the TaqMan-MGB qPCR method established in this study has the advantages of high specificity, high sensitivity, good repeatability, and can be used for rapid and accurate detection of SPPV.
【作者單位】: 中國農(nóng)業(yè)科學院蘭州獸醫(yī)研究所/家畜疫病病原生物學國家重點實驗室;深圳出入境檢驗檢疫局動植物檢驗檢疫技術(shù)中心;
【基金】:國家自然科學基金項目(31201892) 甘肅省國際科技合作專項(1504WKCA055、2016GS08219)
【分類號】:S852.654
本文編號:2186173
[Abstract]:In order to establish a rapid detection method for the (SPPV) gene of sheep pox virus, a pair of specific primers and a MGB probe were designed and synthesized according to the SPPV ORF068 gene in this study. The rapid detection method of SPPV TaqMan-MGB fluorescence quantitative PCR was established by optimizing the conditions. The results showed that this method could be used to detect SPPV-specific, but negative for sheep aphtha virus and porcine blister virus, and the detection limit was 10 copies / 渭 L, and the coefficient of variation of repeated test was less than 3. The above results show that the TaqMan-MGB qPCR method established in this study has the advantages of high specificity, high sensitivity, good repeatability, and can be used for rapid and accurate detection of SPPV.
【作者單位】: 中國農(nóng)業(yè)科學院蘭州獸醫(yī)研究所/家畜疫病病原生物學國家重點實驗室;深圳出入境檢驗檢疫局動植物檢驗檢疫技術(shù)中心;
【基金】:國家自然科學基金項目(31201892) 甘肅省國際科技合作專項(1504WKCA055、2016GS08219)
【分類號】:S852.654
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