四川资阳猪源猪链球菌病病原的分离与鉴定  

Isolation and identification of pathogen related to recent outbreak of pig-human infection in Sichuan province

作　　者：刘华雷[1] 赵永刚[1] 王君玮[1] 赵云玲[1] 李林[1] 郑东霞[1] 孙承英[1] 王珊[1] 刘佩兰[1] 宋翠平[1] 范伟兴[1] 陈义平[1] 王志亮[1] 

机构地区：[1]农业部动物检疫所国家外来动物疫病诊断中心,青岛266032

出　　处：《中华微生物学和免疫学杂志》2005年第12期1006-1010,共5页Chinese Journal of Microbiology and Immunology

摘　　要：目的四川省部分地区发生的不明原因的猪源人畜共患病病原菌的分离与鉴定。方法采用普通RT-PCR和实时荧光PCR相结合的方法对流感和尼帕病毒的RNA进行检测,在Vero细胞和SPF鸡胚同时进行病毒分离,用血清平板对病料进行细菌分离,同时用革兰染色、生化试验、血清凝集试验和PCR进行鉴定,并对分离到的细菌进行药敏试验。结果排除了流感病毒和尼帕病毒感染的可能性。从病料组织中成功分离到3株链球菌,证实为猪链球菌Ⅱ型。通过对其毒力因子进行鉴定,结果发现溶菌酶释放蛋白(MRP)和细胞外蛋白因子(EF)均为阳性。进一步的药敏试验证实:分离菌株对氨苄青霉素、先锋霉素Ⅵ、羧苄青霉素、复方新诺明、头孢呋肟等抗菌药高度敏感。结论病原为猪链球菌Ⅱ型。Objective To isolate and identify the agent responsible for causing a high mortality in the infected people due to the recent zoonotic infection in Sichuan province. Methods Direct reverse transcript PCR and real time PCR were employed to detect the viral RNA of Nipah or Influenza A virus that has been suspected to be in the samples. The virus isolation was carried out with Vero cell and SPF Chicken embryos, and the cultures were subjected to PCR or hemagglutnation（HA ） test for Nipah or Influenza virus after each passage. Bacteria isolation were performed on blood nutrient agar from which the suspected colonies were then selected and tested by agglutination and PCR for Streptococvus suis type Ⅱ. Results The results showed that all the samples and their cultures were negative for influenza virus and Nipah virus in PCR and HA test, suggesting that the possibility of Nipah and Influenza A could be ruled out firm the outbreak. However, three bacterial isolates firm clinical samples were diagnosed as Streptococcus suis type Ⅱ based on Gram staining, biochemical character, agglutination and PCR test. PCR for virulent factor MRP （muramidase-released protein） and EF（ extracellular factor） genes of Streptococcus suis type Ⅱ were all positive in the 3 isolates. Drug sensitivity tests revealed that all the isolates were highly sensitive to ampicillin, cephalothins Ⅵ, carbenicillin, compound sulfamethoxazole and cofuroxime. Conclusion The pig-berne disease outbreaked recently in Sichuan province is supposed to be caused by Streptococcus suis type Ⅱ, which is highly virulent.

关 键 词：猪链球菌Ⅱ型 毒力因子 溶菌酶释放蛋白 细胞外蛋白因子 药敏试验 

分 类 号：S852.611[农业科学—基础兽医学]