副猪嗜血杆菌的分离培养和血清型鉴定

从全国十多个省市送检的疑似患多发浆膜炎与关节炎猪的病料中分离到32株细菌，进行了细菌形态观察、培养特性和生化特性鉴定；根据副猪嗜血杆菌16S rRNA序列设计引物进行PCR扩增，将822bp扩增片段连入T-载体后测序，再与GenBank(M75065)中的序列进行比对，表明其与国外副猪嗜血杆菌菌株16S rRNA序列的同源性为97％以上，确定为副猪嗜血杆菌(Haemophilus parasuis，Hps)。将其中的15株按Kieletein-Rapp-Gabriedson(KRG)琼脂扩散血清分型方法进行血清型鉴定，结果为血清5型3株、血清4型4株、血清13型2株、血清12型2株，另外有4株不能进行分型。该结果表明副猪嗜血杆菌在我国广泛存在。

Isolation and Serovar Identification of Haemophilus parasuis in China

From more than ten provinces and cities in China, 32 strains of Haemophilus parasuis were isolated from swines which characterized by polyserositis and fibrinopurulent polyarthritis, based on morphologic characteristics, cultural characteristics and biochemical characteristics of Haemophilus parasuis . According to the sequence of bacterium's 16S rRNA, a PCR test to diagnose Haemophilus parasuis was developed, and it was used to identify the isolations. The 822 bp amplicon was cloned into a T-vector and sequenced, then the sequence was blasted in the GenBank which showed that its homology is more than 97%. Among these isolates, 15 strains were serotyped in Australia , four serotypes, sv.4(4 strains) , sv.5(3 strains) , sv.12(2 strains) , sv.13(2 strains) were the most frequently isolated stains and 4 stains were classified as nontypable. The results obtained indicate that the distribution of the serotypes in China is very similar to that found by other researchers in Germany, Australia, Canada, Japan, Spain and alike to that found in the United States.