应用不同单克隆抗体和扩增引物快速诊断立克次体的实验研究

The Rapid Detection and Analysis of Rickettsiae with Monoclonal Antibodies to Rickettsial heilongjingii and Primer Pairs for Rickettsial PCR

用不同特异性的黑龙江斑点热立克次体（Rickettsialheilongjiangii）054株单克隆抗体（McAb）或PCR引物检测立克次体的结果表明：群特异性的mcAb2E1和mcAb4G2用微量免疫间接荧光抗体染法（MIIF）法可检测出文中所用各种斑点热立克次体（SFGR）：立氏（R．rickettsii）、康氏（R．conerii）、小蛛（R．akari）、派克（R．parkeri）、西伯利亚（R．sibirica）、澳大利亚（R．australia）和054株立克次体；F7单抗只能检测出R．rickettsii和054株立克次体。PCR扩增结果表明：引物Rt120和Rr120．2048p—1625n分别对恙虫病立克次体（R．tsutsug－amushi）和SFGR扩增出现388bp和523bp扩增带；Rr190．70p—602n和Rr190．4442—5664n两对引物扩增结果相同，即除R．tsutsugamushi外对Q热（C．burnetii）、SFGR、普氏（R．prowazekii）和莫氏（R．mooserii）均扩增阳性，分别为532bp和1222bp电泳带；引物Rpcs．977p—1258n则可扩增上述所有立克次体，电泳带381bp。这样，可在短期内同时检测和初步鉴定不同立克次体。

Detection and analysis of rickeckettsial by monclonal antibodies to Heilonjingii 054 strain of spotted FeverGroup Rickettsiae (SFGR) with micro indirect Immunofluorescence (mllF) and primer pairs for PCR of Rickettsial were reported. The mIIF reaction of mcAb2E and mcAb4G2, were positive with all of the species of SFGRused, included R. rickettsii、 R. sibirica、 R. conorii、 R. pakeri、R. australia and R. heilongjiangii 054 strain; R. rickttsii and 054 strain were detectable with McAbF7,The results of PCR with different primer pairs for Rickettsiaerevealed that 381bp bands appeared with primers RpCS877p-1258n PCR for the followings: R. tsutsugamushi、R. prowazekii、R. mooseri and SFGR. There were 532bp and 1222bp hands with primers R190-70p-602n andRr190. 4442p-5664n for PCR of rickettsiae that were same with those rickettsiae with RpCS primres except R.tsutsugamush. Only apeared the 388bp band of R. tsutsugamushi by PCR with Rt120 and there were 523bp bandsof SFGR by PCR with Rr. 120. 2048p-1625n primer pair. The results above showed that the detection and diagnosis of rickettsiae were possible with these methods.