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目的 了解秋冬型恙虫病流行季节黑线姬鼠、小盾纤恙螨、病人体内恙虫病东方体 (Orientiatsutsuga mushi,Ot)基因型的一致性。方法 采用NestedPCR结合限制性内切酶片段长度多态分析 (RFLP)技术对山东地区恙虫病流行季节病人、野外优势鼠种黑线姬鼠、鼠体外优势螨种小盾纤恙螨标本中Ot-Sta5 6基因型进行分析。结果 应用PCR/RFLP分析了 12株分离自山东秋冬型恙虫病流行季节病人、黑线姬鼠、小盾纤恙螨体内的Ot,2份采自黑线姬鼠体外的小盾纤恙螨匀浆及 10份恙虫病人全血中的Ot-sta5 6目的基因 ,2 4份标本酶切图谱经与已知序列的Ot内切酶图谱相比 ,与日本地方株 -Kawasaki株具有相似的酶切图谱 ,但缺乏HhaI的酶切位点 ;2 4份标本提取的Ot-DNA扩增产物分别经型引物扩增 ,均出现相应分子量的扩增片段 ,皆属日本地方株Kawasaki型。结论 秋冬型恙虫病流行季节病人、野外优势鼠种黑线姬鼠、鼠体外优势螨种小盾纤恙螨体内所分离的恙虫病东方体Sta5 6基因型是一致的 ,并在病人、黑线姬鼠、小盾纤恙螨间构成相互传播关系 ,这对当地恙虫病疫源地的维持有重要作用。
Objective To understand the consistency of genotypes of Orientiatsutsuga mushi (Ot) in autumn and winter epidemic seasons of Apodemus agrarius and Chigger sibiricum. Methods NestedPCR combined with restriction fragment length polymorphism (RFLP) was used to detect the abundance of Ot-2 mRNA in seasonal specimens of patients with tsutsugamushi disease in shandong province and the dominant species of Apodemus agrarius in vitro, Sta5 6 genotypes were analyzed. Results PCR-RFLP analysis of 12 isolates of seasonal autumn, winter and autumn-type tsutsugamushi disease seasons, Apodemus agrarius and Trichocalyptus dorsalis mites were conducted in vitro. Two samples were collected from Apodemus agrarius in vitro Pulsed and stained with Ot-sta5 6 gene from 10 tsutsugamushi’s whole blood samples. Compared with the Ot-endonuclease map of the known sequence, 24 samples showed similar digestion with the Japanese Kawasaki strain Map, but the lack of HhaI restriction sites; 24 samples extracted Ot-DNA amplification products were amplified by primer, there are corresponding molecular weight amplified fragments, are Japan’s local strains Kawasaki type. Conclusions The seasonal Sta5 6 genotype of Orientia tsutsugamushi isolated from chickens infected with autumn and winter tsutsugamushi disease, the dominant species of black-spotted Apodemus agrarius and the predominant species of mite in the outside of the body are consistent with each other. Apodemus, a small shield fiber mite mites constitute a mutual transmission relationship, which is the local scrub typhus epidemic maintain an important role.