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本文PCR技术的引物是参考恙虫病立克次体Karp株Sta 58序列设计合成的一对DNA引物.以恙虫病立克次体DNA为模板,成功扩增长约1kb的DNA片段.该对引物首次应用于人工成虫腹内接种羔虫病立克次体(人工接种法)和幼虫叮咬恙虫病鼠(叮咬病鼠法)的我国主要恙虫病媒介地里纤羔螨的研究.PCR检测人工接种法成虫的不同时期和经卵传递的子代立克次体DNA均获满意结果,立克次体在恙螨体内生长持续360天及产卵孵出的第4代幼虫均呈阳性.叮咬病鼠法立克次体在恙螨亲代饱食蚴、若蛹、若虫、成虫及第2代子产代幼虫,PCR检测均呈阳性.结果显示PCR技术检测恙螨体内恙虫病立克次体的特异性和敏感性高;体外基因扩增检测恙螨体内及鼠宿主体内立克次体是流行病学调查的新的敏感方法.本法为恙虫病分子体流行病学调查提供了新的应用技术,亦为临床病人诊断提供了敏感方法.
The primers of this PCR technique are a pair of DNA primers designed and synthesized with reference to the Sta 58 sequence of Rickettsia tsutsugamushi Karp Strain. The DNA fragment of about 1 kb in length was successfully amplified by using the Rickettsia tsutsugamushi Rickettsia DNA as a template. The first application of artificial insects intra-abdominal vaccination of rickettsia rickettsia (artificial inoculation method) and larvae bite tsutsugamushi disease (bite disease rat) in China’s major scrub typhus vectors fiber Bombyx mori. PCR detection of artificial inoculation The results showed that rickettsia grew well in chigger mites for 360 days and oviposit 4th generation larvae were positive. Rickettsia rickettsiae could secrete larvae in chigger mites progeny, and all the PCR products were positive if they were pupae, nymphs, adults and second-generation progeny larvae. The results showed that PCR detection of Rickettsia tsutsugamushi Specificity and sensitivity.It is a new and sensitive method for epidemiological investigation of chigger mites in vivo and in vivo in vivo by gene amplification.This method provides a new application for molecular epidemiology of scrub typhus Technology, but also for the clinical diagnosis of patients with sensitive methods.