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为了快速检测临床上副猪嗜血杆菌(Haemophilus parasuis,Hps)感染,根据Gen Bank中Hps的16S rRNA基因保守区序列,设计1对特异性引物,利用PCR方法检测Hps,并进行特异性、敏感性及临床检测。结果显示:PCR反应选择58℃为最适退火温度,0.5μL(20μmol/L)为最适引物量,模板浓度为7.2×10~(-6)μg/mL时即可扩增出814 bp特异性目的片段,与细菌分离结果一致。结果表明:该方法敏感、快速、特异,可用于临床上Hps感染的快速检测。
In order to rapidly detect the infection of Haemophilus parasuis (Hps) in clinic, one pair of specific primers was designed according to the sequence of 16S rRNA gene of Hps in Gen Bank. Hps was detected by PCR method and was specific and sensitive Sexual and clinical testing. The results showed that the best annealing temperature was 58 ℃ for PCR reaction, and the optimum amount of primer was 0.5μL (20μmol / L). When the template concentration was 7.2 × 10 ~ (-6) μg / mL, Sexual fragments, consistent with the results of bacterial isolation. The results show that this method is sensitive, rapid and specific and can be used for the rapid detection of Hps infection in clinic.