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中文摘要：
根据GenBank中登录的猪肺炎支原体P36（L-乳酸脱氢酶）全基因序列，设计合成了2对引物，建立套式PCR方法。运用该方法对猪肺炎支原体不同菌株培养物进行了扩增，并对经肺内免疫猪支原体肺炎活疫苗后支气管肺泡灌洗液进行了检测。结果表明，不同菌种均能扩增出427bp的目的条带，而其他病原菌未出现特异性扩增条带。建立的套式PCR方法检测猪肺炎支原体的最低检测限度为10个CCU（颜色变化单位）。支气管肺泡灌洗液检测结果表明，经肺内免疫的猪支气管肺泡灌洗液扩增结果均为阳性。
英文摘要：
Two pairs of primers were designed according to the sequence of P36 （L-lactate dehydrogenase） gene of Mycoplasma hyopneumoniae published in GenBank, and a nested PCR assay was established for the detection of M. hyopneu- moniae. Different M. hyopneumoniae strains and the bronchoalveolar lavage fluid of intrapulmonic immunization with attenua- ted M. hyopneumoniae vaccine were detected with nested PCR. A specific 427 bp fragment was amplified from different M. hyopneumoniae strains by the developed nested PCR. Non-M. hyopneumoniae did not show specific amplified fragment. The nested PCR had high sensitivity to for detection of 10 CCU M. hyopneumoniae. The results of the bronchoalveolar lavage fluid detected by nested PCR showed that the amplified results was positive by intrapulmonic immunization.
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