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首页> 外文期刊>African Journal of Biotechnology >Improvement of specific polymerase chain reaction (PCR) for the identification of Mycoplasma capricolum subsp. capripneumoniae
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Improvement of specific polymerase chain reaction (PCR) for the identification of Mycoplasma capricolum subsp. capripneumoniae

机译：特异性聚合酶链反应（PCR）的改进用于鉴定支原体亚种。肺炎克雷伯菌

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A?polymerase chain reaction (PCR)?test for the identification of?Mycoplasma capricolum?subsp.?capripneumoniae?(Mccp) was optimized using the H2 gene sequences of M1601. The test was evaluated on 20 strains including six representative strains of the?Mycoplasma mycoides?cluster as well as 13 field isolates from China,?Pasteurella multocida and Mannheimia haemolytica. To obtain a test that was specific for Mccp, the PCR produced an amplicon of approximately 680 bp only with the Mccp strains. The specificity of the present PCR was found to be 100% specific for Mccp. The sensitivity of the PCR showed that it could detect a minimum of 0.75 ng of purified DNA. For further evaluation, clinical samples tested by PCR included lung and liquor pleurae from 14 goats artificially infected with Mccp. The PCR was positive in all the clinical samples both in the undiluted and 50-fold dilutions of extracted DNA except for a sample of liquor pleurae. However, the latter showed weak bands. An epidemiological survey was also performed using the constructed PCR methods and results indicated that 36 of 61 of these samples including tissue and percolate from Western China were considered positive. The coincidence rate was 82% compared with the performance of a PCR test earlier published. The present PCR represents a rapid and reliable method for genetically based identification of Mccp. The specificity of the test makes it suitable for detection of Mccp in clinical samples.
机译：利用M1601的H2基因序列优化了用于鉴定“支原体支原体”亚种“ capripneumoniae”（Mccp）的“聚合酶链反应（PCR）”测试。该测试对20株菌株进行了评估，其中包括6株代表性支原体“ Mycoplasma mycoides”集群以及13株来自中国的多形巴斯德氏菌和溶血曼海姆氏菌。为了获得对Mccp特异的测试，仅使用Mccp菌株，PCR产生了大约680 bp的扩增子。发现本PCR的特异性对Mccp是100％特异性的。 PCR的敏感性表明，它可以检测到至少0.75 ng的纯化DNA。为了进一步评估，通过PCR测试的临床样本包括来自14只人工感染了Mccp的山羊的肺和胸膜胸膜。在未稀释的和稀释50倍的提取DNA中，所有临床样品中的PCR均为阳性，但胸膜白酒样品除外。但是，后者显示出较弱的波段。还使用构建的PCR方法进行了流行病学调查，结果表明，在这些样本中，有61种中的36种（包括来自中国西部的组织和渗滤液）被认为是阳性。与早先发表的PCR测试相比，符合率为82％。本PCR代表了一种快速可靠的基于基因的Mccp鉴定方法。该测试的特异性使其适合检测临床样品中的Mccp。

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