应用广谱引物PCR法检测立克次体的研究

Detection of Rickettsia by Extensive Primer PCR Method

本文选用普氏立克次体柠檬酸合成酶基因做为靶序列合成一对引物（ＲＰＣＳ８７７ｐ和ＲＰＣＳ１２５８ｎ）。用该对引物对不同种类立克次体（包括：加拿大；普氏；莫氏；Ｑ热；恙虫病Ｇｉｌｌｉａｍ株、Ｋａｔｏ株、Ｋａｒｐ株；澳大利亚；立氏；康氏；小蛛；西伯利亚２４６、西伯利亚２３２；精河；内０１；内Ｗ８８；黑８４；黑０５４）及大肠杆菌ＨＢ１０１，ＤＮＡ进行扩增。同时，选用三种不同方法处理模板进行扩增结果的比较。结果表明：该引物可扩增除恙虫病以外的所有立克次体，而大肠杆菌、λＤＮＡ不能被扩增。采用简单煮沸法处理模板，对含立克次体的材料进行扩增是可行的。

Polymerase chain reaction(PCR)is a potentially effective method for detecting Rickettsial Infections.For this reason,an assay for diagnosis and epidemiological investigations of rickettsij,was developed Rickettsia prowazekii citrate synthase gene was selected as target sequence and a pair primer(Rpcs877p and Rpcs1258n) was synthesiged.It was amplified that included international strain(R. Canada;R. Prowazekii;R. Mooseri;C.Burneti;R.tsutsugamushi;R. Rickettsi;R. sibirica;R. Conori;R.Australis;R. Akari; R.parkeri)、chinese isolates (strain jinghe;heilongjiang(strain 054.strain 84);inner mongolia (strain 01;strain w88)、E. coliHB101 and λDNA.The results showed that the primer could amplified various rickettsii except R. tsutsugamushi、E.coji HB101 and λDNA.It was possible that simple boiling method could prepare template,The PCR method can be used in epidemiological screen.