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中国分离出的三株新的斑点热群立克次体的基因型鉴定

Genotypic identification of three new strains of spotted fever group rickettsiae isolated in China.

作者信息

Zhang J Z, Fan M Y, Bi D Z, Cui W F, Han Y F

机构信息

Department of Rickettsiology, Chinese Academy of Preventive Medicine, Beijing, P.R. of China.

出版信息

Acta Virol. 1996 Sep;40(4):215-9.

PMID:9014012
Abstract

Polymerase chain reaction (PCR) and restriction endonuclease fragment length polymorphism (RFLP) analysis were used to characterize the genotypic diversity of three isolates of spotted fever group (SFG) rickettsiae isolated from ticks in China. A primer pair designed from DNA sequence encoding 190 K protein antigen of R. rickettsii and genomic DNAs obtained from the isolates were used in PCR. The PCR products were cleaved with restriction endonucleases PstI and RsaI, and the digestion patterns were analyzed by polyacrylamide gel electrophoresis (PAGE) and compared with those of all known species and strains of SFG rickettsiae. The results showed that three isolates had the same PCR products as the other SFG rickettsiae under comparison. HL-93 strain, isolated from Hemophysalis concinna ticks collected in Hulin County, Heilongjiang Province, had unique PstI digestion pattern among SFG rickettsiae; strains BJ-93 and 053, isolated from Dermacentor sinicus and Haemaphysalis concinna ticks collected in Changping County, Beijing City, and Suifenhe City, Heilongjiang Province, respectively, had the same PstI and RsaI digestion patterns as strains R. sibirica 246, BJ-90 and IMTO-85. The present study demonstrated that the BJ-93 and 053 strains were genotypically identical with R. sibirica and the HL-93 strain was genotypically unique among SFG rickettsiae.

摘要

采用聚合酶链反应(PCR)和限制性内切酶片段长度多态性(RFLP)分析方法,对从中国蜱虫中分离出的3株斑点热群(SFG)立克次体的基因型多样性进行了鉴定。根据立氏立克次体190K蛋白抗原编码DNA序列设计引物对,并将其与从分离株中获得的基因组DNA用于PCR。PCR产物用限制性内切酶PstI和RsaI进行切割,消化模式通过聚丙烯酰胺凝胶电泳(PAGE)进行分析,并与所有已知的SFG立克次体物种和菌株进行比较。结果显示,3株分离株与其他被比较的SFG立克次体具有相同的PCR产物。从黑龙江省虎林县采集的嗜群血蜱中分离出的HL-93菌株,在SFG立克次体中具有独特的PstI消化模式;分别从北京市昌平区和黑龙江省绥芬河市采集的中华革蜱和嗜群血蜱中分离出的BJ-93和053菌株,其PstI和RsaI消化模式与西伯利亚立克次体246、BJ-90和IMTO-85菌株相同。本研究表明,BJ-93和053菌株在基因型上与西伯利亚立克次体相同,而HL-93菌株在SFG立克次体中基因型独特。

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