【摘要】 通过反应条件的优化、特异性和敏感性的测定,建立了一种快速、简便、准确的PCR方法,用于检测副猪嗜血杆菌(HPs)。PCR扩增片段大小为822bp,最小检测量为1080个HPs。与大肠杆菌、沙门氏菌、金黄色葡萄球菌和链球菌无特异性反应。用该法检测人工感染猪的肺脏、心脏和肾脏抽提的DNA,结果为阳性;检测肝脏、脾脏、脑组织样品抽提的DNA,结果为阴性。用该法检测人工感染发病仔猪及自然感染仔猪肺组织,并结合细菌分离培养,证明以肺脏组织为病料,通过细菌分离培养,再用PCR法鉴定分离菌体,可有效地用于HPs的诊断。检测110份来自发病猪场的患病仔猪肺脏病料,其中有53份为阳性,表明HPs所致发的Glasser′s病已在我国部分地区发生和流行。更多还原

【Abstract】 A PCR assay was developed to detect Haemophilus parasuis quickly and accurately.The size of the amplified PCR product was 822 bp.The sequence of the PCR product had the characterization of the 16S rRNA of H.parasuis.The minimum dose of H.parasuis was 1 080 bacteria.Four H.parasui isolates could be detected using the PCR test.But no amplification was observed when other 4 bacterial species isolated from swine tissue were detected.Six clinical samples,from 3 piglets experimentally infected with H.parasuis isolates and 3 naturally infected piglets,were tested by PCR.The results showed that the PCR assay could be used to detect H.parasuis,associated with bacteria isolate for lung sample.When the assay was used to detect the sick piglets lung sample from 110 pig farm,the results showed that the 53 samples of which were positive to H.parasuis.It indicated that the PCR assay could be used to detect H.parasuis,and that its resultant disease,the Glasser′s disease has been existed and prevails epidemically in some region of China.更多还原