副猪嗜血杆菌分离鉴定与PCR检测方法的建立及应用  

Isolation and Identification of Haemophilus Parasuis and Establishment of PCR Detection and Application

作　　者：林雪玲[1] 黄耿森[1] 冼琼珍[1] 黄良宗[1] 顾万军[1] 

机构地区：[1]佛山科学技术学院生命科学学院,佛山528231

出　　处：《吉林农业大学学报》2006年第3期321-324,共4页Journal of Jilin Agricultural University

基　　金：广东省自然科学基金资助项目(64064)

摘　　要：从养猪场送检的猪肺病料中分离到3株疑似副猪嗜血杆菌(HPS)可疑菌株。经形态镜检、染色特性、生化和部分生物学特性试验,初步鉴定为HPS。在细菌学鉴定基础上,设计合成3对引物,分别以3株分离菌株的DNA为模板进行PCR扩增,结果均扩增出822 bp8、24 bp和1.9 kb的预期特异性条带。以HPS GD株作为参考阳性菌株,以P1、P2为引物,设计优化PCR反应程序,建立HPS PCR检测方法,并应用于临床病料检测。结果表明,所建立的HPS PCR检测方法特异与敏感性高、适用性强,可应用于HPS感染的诊断和检测。3 stains of Haemophilus Parasuis（HPS） were isolated from swine lungs which may be infected with HPS in Guangdong and Guangxi provinces. The strains are identified based on morphologie, cultural and biochemical characteristics of HPS. With 3 pairs of primers according to the sequences of the HPS＇s 16s rRNA and other genes, the PCR test was developed and PCR products including 822 bp, 824 bp and 1.9 kb fragments were observed with DNA of the strains of HPS. The GD stain was as positive reference strain. PCR detecting method is estabhshed with the fragment of HPS＇s 16s rRNA. The experiment resuits show that the PCR detecting method is specific and sensitive and can be used for detecting and identifying HPS infection.

关 键 词：副猪嗜血杆菌 分离鉴定 PCR 

分 类 号：S852.612[农业科学—基础兽医学]