【摘要】：目的建立巴斯德桿菌的CODEHOP PCR快速檢測(cè)方法,為實(shí)驗(yàn)動(dòng)物的呼吸道細(xì)菌的控制提供參考。方法應(yīng)用CODEHOP在線簡(jiǎn)并引物設(shè)計(jì)工具,比對(duì)Genbank中13株巴斯德桿菌的RNA聚合酶β亞基(rpo B)氨基酸序列設(shè)計(jì)簡(jiǎn)并引物。對(duì)建立CODEHOP PCR方法用21株參考菌株進(jìn)行特異性和敏感性評(píng)價(jià),并應(yīng)用于實(shí)驗(yàn)動(dòng)物中的巴斯德桿菌檢測(cè)。結(jié)果簡(jiǎn)并引物Past F6/PastR5擴(kuò)增標(biāo)準(zhǔn)菌株的目的片段為200 bp左右。能夠區(qū)分受試的巴斯德桿菌和主要的實(shí)驗(yàn)動(dòng)物呼吸道病原菌。敏感性為0.2 pg/μL~2 pg/μL。在受試的609只實(shí)驗(yàn)動(dòng)物中呼吸道樣品中檢測(cè)出巴斯德桿菌陽(yáng)性率為19.1%。樣品陽(yáng)性片段經(jīng)測(cè)序驗(yàn)證,準(zhǔn)確率為100%。結(jié)論所建立的方法具有良好的特異性和敏感性,可用于動(dòng)物樣品中巴斯德桿菌的檢測(cè)。
[Abstract]:Objective to establish a rapid CODEHOP PCR method for the detection of Pasteurella vulgaris and to provide reference for the control of respiratory tract bacteria in laboratory animals. Methods the degenerate primers of RNA polymerase 尾 subunit (rpo B) of 13 Pasteurella strains in Genbank were designed by using CODEHOP online degenerate primer design tool. The CODEHOP PCR method was used to evaluate the specificity and sensitivity of 21 reference strains and was applied to the detection of Pasteurella in laboratory animals. Results the target fragment of standard strain amplified by degenerate primer Past F6/PastR5 was about 200 bp. Pasteurella can be distinguished from the main laboratory animals respiratory pathogens. The sensitivity was 0.2 pg/ 渭 L ~ (2) pg/ 渭 L. The positive rate of Pasteurella was 19.1 in the respiratory tract samples of 609 experimental animals. The positive fragment was confirmed by sequencing and the accuracy was 100%. Conclusion the method has good specificity and sensitivity and can be used for the detection of Pasteurella in animal samples.
【作者單位】： 中國(guó)食品藥品檢定研究院實(shí)驗(yàn)動(dòng)物資源研究所;中國(guó)醫(yī)學(xué)科學(xué)院/北京協(xié)和醫(yī)學(xué)院醫(yī)學(xué)生物學(xué)研究所樹(shù)，

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