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目的 用分子生物学技术鉴定南澎列岛恙虫病东方体及其分型。方法 以巢式聚合酶链反应 (NPCR)检测南澎列岛恙虫病东方体 8个分离株的 5 6kD蛋白基因片段 ,阳性产物进行限制性片段长度多态性分析 (RFLP)。选择其中 4株的PCR产物克隆进 pGEM -T载体并且测序 ,测序结果在国际互联网作多序列比较和进化树分析。 结果 7株分离株扩增出5 0 0bp目的片段 ,RFLP分析表明南澎列岛存在三种不同的恙虫病东方体。从 4个样品获得 7个克隆 ,序列分析证实 2个克隆(DY1,DY2 1)与Yonchon同源性 99% ,3个克隆 (DY2 2 ,NY12 ,NS2 )与Karp株同源性 92 % ,2个克隆 (NY11,NS1)与Kato株同源性 98%。结论 南澎列岛存在Karp ,Kato ,Yonchon三种型别的恙虫病东方体
Objective To identify the Oriental body of tsutsugamushi disease and its classification in Nanpeizan Island by molecular biology techniques. Methods The 5 6 kD gene fragment of 8 isolates of Orientia tsutsugamushi from southern Taiwan Penaeus monocytogenes was detected by nested polymerase chain reaction (NPCR). The positive products were analyzed by restriction fragment length polymorphism (RFLP). Four of the selected PCR products were cloned into pGEM-T vector and sequenced. The sequencing results were compared on the Internet for multiple sequence comparison and phylogenetic tree analysis. Results A total of 500 bp fragments were amplified from 7 isolates. RFLP analysis showed that there were three different oriental strains of tsutsugamushi in South Peninsula. Seven clones were obtained from four samples. Sequence analysis confirmed 99% homology between Yonchon and two clones (DY1, DY2 1), 92% homology between Karp and two strains (DY2 2, NY12 and NS2) One clone (NY11, NS1) had 98% homology with the Kato strain. Conclusion There are three types of tsutsugamushi bodies such as Karp, Kato and Yonchon in Nanpeng Islands