斑点热群立克次体新种──虎林－93株的分离和鉴定

Isolation and Identification of New Species of Spotted Fever Group Rickettsiae

本文用鸡胚卵黄囊培养法及实验动物感染法从黑龙江省虎林县境内采集的嗜群血蜱中分离到了一株立克次体。命名为ＨＬ－９３株立克次体。经形态学及血清学试验证实为斑点热群立克次体。分别用微量免疫荧光法、十二烷基硫酸钠－聚丙烯酰胺凝胶不连续电泳法、单克隆抗体免疫酶斑点法、单克隆和多克隆抗体免疫印迹法、聚合酶链反应和限制性片段长度多态性分析法对该分离株进行鉴定并同已知国际标准株、国际及国内参考株进行比较。结果表明；ＨＬ－９３株立克次体无论在抗原性多肽上，还是在基因水平上在斑点热群立克次体中都是独特的。根据目前立克次体分类法，ＨＬ－９３株立克次体可以考虑是斑点热群立克次体的新种。

One strain of rickettsiae was isolated from Haemaphysalis concinna collected from Hulin county in Heilongjiang province in 1993 by using embryonated hens eggs and experimental animals and it was proved to be the member of spotted fever group rickettsiae (SFGR) by the methods of morphology and serology, and named as HL-93 strain after the name of the place and the year which it was isolated. The isolate of spotted fever group rickettsiae was compared with known species and strains of SFGR by Microimmunofluorescence assay, DNA polymerase chain reaction, DNA polymerase chain reaction followed by restriction endonuclease fragment length polymorphism analysis, Sodium dodecylsulphate-polyacrylamide gel electrophoresis, protein immunoblotting with polyclonal and monoclonal antibodies, dot immunobinding assay. The results demonstrated that HL-93 strain was antigenically and genotypically unique among spotted fever group rickettsiae. We feel that HL-93 strain should be considered as a new member of spotted fever group rickettsiae on the basis of current rickettsiae taxonomy.