PCR技术在鼠肺支原体检测中应用的初步研究

目的　探讨PCR技术在鼠肺支原体检测中的应用,希望能建立一种可行、快速、敏感的检测方法。方法　使用支原体通用引物及鼠肺支原体特异性引物对14 份大鼠喉气管拭子洗液和拭子支原体培养液进行PCR扩增,2 % 琼脂糖电泳鉴定。另设M53 和ATCC19612 二株标准鼠肺支原体菌株作阳性对照。结果　通用引物对大鼠喉气管拭子洗液检出率8/14 ,拭子支原体培养液检出率14/14,鼠肺支原体特异引物PCR扩增对大鼠喉气管拭子洗液检出率0/14 ,拭子支原体培养液3/14。通用引物扩增M53 和ATCC19612 二株标准株均呈现阳性,而鼠肺支原体特异引物扩增M53 和ATCC19612,只有M53 呈现阳性。结论　PCR通用引物检测比普通分离培养省时省力,而我们采用国外某学者认为对鼠肺支原体有特异性的引物,是否可用于鼠肺支原体的特异性PCR 检查仍需进一步探讨。

The Pilot Study of PCR Assay for the Detection of Mycoplasma Pulmonis in Infected Rats

To investigate the PCR (polymerase chain reaction, PCR) assay for the detection of Mycoplasma pulmonis in infected rats and try to set up a feasible, quick and sensitive method for the diagnosis. Method 14 samples of the supernatants of throat swab samples and the cultural supernatants of throat swab from infected rats are performed PCR analysis using animal mycoplasms routine primers and M. Pulmonis species-specific primers respectively. The PCR products are confirmed with 2% agarose gel electrophoretic analysis. The M 53 and ATCC19612 standard strain of Mycoplasma pulmonis are used as positive control. Results Using animal mycoplasma routine primers, 8/14 of the supernatants of throat swab samples are positive in the PCR assay whereas 14/14 of the cultural supernatants from throat swab specimen are positive. With M.Pulmonis species-specific primers, the supernatants of throat swab samples are negative in the PCR analysis but 3/14 of the cultural supernatants from throat swab samples are positive. The mycoplasma routine primers can detect the M 53 and ATCC19612 standard strain, on the other hand, M.pulmonis species-specific can only detect M 53 but ATCC19612 strain. Conclusions The PCR assay using mycoplasma routine primers is superior to isolated culture for detecting M. Pilmonis, which could save time and labor. Nevertheless, whether the M.Pilmonis species-specific primers which the foreign researchers suggested could be used to specially detect the M.Pulmonis in the infected rats, need to be further investigated.