血液直接PCR方法在猪嗜血支原体检测中的应用

Establishment and application of direct PCR of blood for detecting Mycoplasma suis

为临床上能快速准确检测猪嗜血支原体感染,建立了一种检测猪嗜血支原体的血液直接PCR方法。该方法能特异性地扩增猪嗜血支原体16SrRNA基因中的一段396bp序列,而对猪繁殖与呼吸综合征病毒、猪圆环病毒2型、猪细小病毒、猪伪狂犬病病毒、大肠杆菌、金黄色葡萄球菌等没有扩增到目的条带;该方法能检测到每微升抗凝血中猪嗜血支原体的最低拷贝数为5.52×101 copies,与常规PCR相当;应用血液直接PCR和常规PCR对100份抗凝血样进行检测,阳性检出率分别为65%和69%,差异不显著(P>0.05)。结果表明,血液直接PCR方法具有良好的特异性、较高的灵敏度和较快的检出速度,适用于临床上对猪嗜血支原体感染进行快速确诊和流行病学调查。

In order to detect Mycoplasma suis rapidly in clinical,the direct PCR of blood was established and used.The method could selectively amplify a 396 bp DNA fragment of the 16 SrRNA gene of Mycoplasma suis.No PCR products were amplified from the samples of PRRSV,PCV2,PPV,PRV,E.coli,S.aureus.The lowest detectable limit of the method was 5.52×101copies of Mycoplasma suis per microliter anticoagulated blood,which was equivalent to conventional PCR.Using the direct PCR of blood and conventional PCR,100 anticoagulated blood samples from clinical were detected,positive rate of them was 65%and 69%separately,the difference was not significant at P〉0.05.The results showed that the direct PCR of blood had good specificity,high sensitivity,fast detection speed,and was suitable for the quick diagnosis of Mycoplasma suis infection in clinical and epidemiological investigation.