聚合酶链反应用于恙虫病立克次体检测和分型的研究

Studies on Detecting and Typing of Rickettsia Tsutsugamushi by polymerase Chain Rea ction

本文报道聚合酶结反应用于恙虫病立克次体（Rt）检测和分型研究结果，以构建的Rt外膜主要蛋白56kDa型特异抗原（tsa56kDa）基因编码区的群、型特异引物，采用1步法PCR分别对Gilliam，Karp，Kato，Kawasaki和Kuroki5株标准株Rt，江苏地区5份Rt阳性标本，斑点热和莫氏立次体以及5份正常输血员全血标本进行检测和分型。研究结果证明所构建的Rt群、型引物具有Rt的特异性，型特异引物在Rt的型与型之间无交叉扩增。因此，该研究对恙虫病的诊断、流行病学调查以及Rt的型别鉴定具有实用价值。通过对江苏省东台、海安、南京地区5株Rt的型别鉴定，再次证明江苏地区Rt流行株为我国首次从分子水平发现的一个新型Rt．而且是江苏地区流行的主要优势株，与日本Kawasaki型相似。在调查中未发现其它株型。

Institute qf Military Medicine, Nanjing Command PLA, Nanjing 210002One-step polymerase chain reaction (PCR) method was used for detecting and typing Rickettsia tsutsugamushi (Rt). The primers of the group and type were derived from the type-specific antigen (Tsa) gene DNA sequences (encodes the 56kDa antigen) of standard Rt, Gilliam, Karp, Kato, Kawasaki. Kuroki and Shimokoshi strains. These primers served to produce rickettsia-specific products in the amplification of templated DNA prepared from 6 strains standard Rt and 5 Positive Rt samples selected from Jiangsu Provinde 1No products in it fromRickettsia sPotted fever,Rickettsia mooseri and 5 blood donors. The typing primers pairs did not amplify crossly in types. The results indicated that these primer pairs were of the characteristic of Rt .it was useful for diagnosis,typing and investigating in epidemiology.The typing results indicated akain that the Rt discover belonged to a new serotype and dominant strain in epidemic in Jiangsu. This Rt was similar to Kawasaki strain of Japan. No another Rt strains were found in this study.