本文選題：多殺巴氏桿菌 + 支氣管鮑特桿菌　； 參考：《中國(guó)比較醫(yī)學(xué)雜志》2017年08期

【摘要】：目的建立快速檢測(cè)多殺巴氏桿菌、支氣管鮑特桿菌、支原體和肺炎克雷伯桿菌4種實(shí)驗(yàn)動(dòng)物病原體的多重PCR方法。方法根據(jù)Gen Bank基因設(shè)計(jì)出特異性引物;經(jīng)過多重PCR的優(yōu)化,特異性和敏感性的檢測(cè),建立多重PCR體系。應(yīng)用該P(yáng)CR體系檢測(cè)人工感染樣本和實(shí)驗(yàn)動(dòng)物的氣管分泌物,并與傳統(tǒng)方法做對(duì)比。結(jié)果多重PCR擴(kuò)增出多殺巴氏桿菌(356 bp)、支氣管鮑特桿菌(237 bp)、支原體(266 bp)和肺炎克雷伯桿菌(142 bp)的目的條帶。肺炎克雷伯桿菌敏感性為10 pg,多殺巴氏桿菌、支氣管鮑特桿菌、支原體敏感性為1 pg,特異性檢測(cè)未從其他病原菌中檢測(cè)出目的條帶。應(yīng)用建立的多重PCR體系檢測(cè)人工感染樣本的不同組合,45只實(shí)驗(yàn)動(dòng)物氣管檢測(cè)出15只多殺巴氏桿菌陽性,9只肺支原體陽性,但傳統(tǒng)培養(yǎng)方法和血清學(xué)方法未檢測(cè)出陽性標(biāo)本。結(jié)論本文建立的多重PCR方法操作簡(jiǎn)單、快速、特異性強(qiáng)、靈敏度高,能夠?qū)崿F(xiàn)對(duì)多殺巴氏桿菌、支氣管鮑特桿菌、支原體和肺炎克雷伯桿菌4種實(shí)驗(yàn)動(dòng)物病原體的快速檢測(cè)。
[Abstract]:Objective to establish a multiplex PCR method for rapid detection of four pathogens of Pasteurella multocida, Bacillus bronchus, Mycoplasma pneumoniae and Klebsiella pneumoniae. Methods specific primers were designed according to the Gen Bank gene, and the multiplex PCR system was established by the optimization of multiple PCR and the detection of specificity and sensitivity. The PCR system was used to detect trachea secretions from artificially infected samples and experimental animals, and compared with traditional methods. Results the target bands of Pasteurella multocida, Pasteurella pneumoniae, Mycoplasma pneumoniae and Klebsiella pneumoniae were amplified by multiplex PCR. The sensitivity of Klebsiella pneumoniae was 10 PG, that of Pasteurella multocida, Bacillus bronchus and Mycoplasma was 1 PG. Using the multiplex PCR system to detect the different combinations of artificial infection samples of 45 experimental animals, 15 of them were positive for Pasteurella multocida in the trachea, 9 were positive for Mycoplasma multogenes, but no positive specimens were detected by traditional culture methods and serological methods. Conclusion the multiplex PCR method established in this paper is simple, rapid, specific and sensitive. It can be used to detect the pathogens of Pasteurella multocida, Pasteurella bronchus, Mycoplasma pneumoniae and Klebsiella pneumoniae.
【作者單位】： 河北省實(shí)驗(yàn)動(dòng)物重點(diǎn)實(shí)驗(yàn)室河北醫(yī)科大學(xué)實(shí)驗(yàn)動(dòng)物學(xué)部;柏鄉(xiāng)縣中心醫(yī)院;河北醫(yī)科大學(xué)第四醫(yī)院;
【基金】：國(guó)家科技支撐計(jì)劃(2015BAI07B02-05)
【分類號(hào)】：Q78;R-33

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