ORFV和Mo雙重PCR檢測(cè)方法的建立及應(yīng)用
[Abstract]:Sheep infectious pustular virus (Orf virus,ORFV) is the pathogen of infectious pustular disease (Contagious ecthyma,CE), and mycoplasma pneumoniae (Mycoplasma ovipneumoniae,Mo) is the main pathogen of (Mycoplasma pneumonia of goats and sheep,MPGS. In recent years, the mixed infection of these two pathogens is increasing day by day. At present, a single PCR method is used to detect the two pathogens. In order to establish a double PCR method for the simultaneous detection of ORFV nucleic acid and Mo nucleic acid from the samples, the sequence of the main membrane protein (major envelope protein B2L gene of ORFV and the membrane protein P80 of Mo (membrane protein P80 gene was studied. Two pairs of specific primers were designed and synthesized. The results showed that the specific fragments of 402 bp of ORFV and 700 bp of Mo could be amplified simultaneously by this method, but the DNA templates of other common pathogens were not amplified, showing good specificity. The minimum detection levels for ORFV and Mo were 1.1 脳 10~3copies/ 渭 L and 3.47 脳 10~3copies/ 渭 L, respectively, which were 10 times more sensitive than single PCR. The (Capra hircus) clinical samples of 883 sick goats from some sheep farms in Fujian Province were detected. The positive rate of ORFV was 33. 7% (33. 7%) and 33 (39. 8%) by double PCR. 9 samples (10.8%) were positive for both ORFV and Mo, and the results were consistent with those of single ORFV and Mo PCR. The dual PCR method established in this study can be used for the clinical detection of ORFV and Mo, and provides an effective method for rapid differential diagnosis and epidemiological investigation of CE and MPGS.
【作者單位】： 福建省農(nóng)業(yè)科學(xué)院畜牧獸醫(yī)研究所;
【基金】：福建省農(nóng)業(yè)科學(xué)院科技創(chuàng)新項(xiàng)目(No.PC2017-9) 福建省科技創(chuàng)新平臺(tái)建設(shè)項(xiàng)目(No.2014N2003-5) 國(guó)家重點(diǎn)研發(fā)計(jì)劃(No.2016YFD0500906) 福建省農(nóng)業(yè)科學(xué)院科技創(chuàng)新團(tuán)隊(duì)PI項(xiàng)目(No.2016PI-7)
【分類(lèi)號(hào)】：S852.65
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