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目的鉴定山东地区恙虫病东方体分离株的基因型别,分析其核苷酸序列。方法以日本、韩国常用的恙虫病东方体分型引物,用巢式PCR技术检测恙虫病东方体分离株的相对分子质量(Mr)56×103蛋白基因片段,对群引物扩增的PCR产物纯化回收后测序,并行序列分析。结果Gilliam、Karp、Kato、山东分离株用群引物均扩增出481-507 bp的目的片段,Gilliam、Karp、Kato经相应的型引物扩增出了目的片段。山东分离株仅Kawasaki型的分型引物扩增出目的条带,序列分析证实山东分离株与Kawasaki型相似。结论在国内首次扩增出Gilliam、Karp、Kato三型标准株,山东地区恙虫病东方体分离株为Kawasaki型;山东地区恙虫病东方体与日本的Kawasaki型相似,两者有较近的亲缘关系。
Objective To identify the genotypes of oriental isolate of tsutsugamushi in Shandong Province and analyze its nucleotide sequence. Methods The molecular weight (Mr) 56 × 103 of M. tsutsugamushi isolated from oriental isolates of Orientia tsutsugamushi was detected by nested PCR using the common orienting primers of Orientia tsutsugamushi in Japan and South Korea. The amplified PCR products were purified Recycling after sequencing, parallel sequence analysis. Results The 481-507 bp fragment was amplified by Gilliam, Karp, Kato and Shandong isolates. The target fragments were amplified by Gilliam, Karp and Kato respectively. Only Kawasaki-type genotyping primers of Shandong isolates amplified the target bands, and sequence analysis confirmed that Shandong isolates were similar to Kawasaki type. CONCLUSIONS Three types of standard strains of Gilliam, Karp and Kato were amplified for the first time in China. The Kawasaki type isolates of tsutsugamushi in Shandong province were Kawasaki type. The Kawasaki type of tsutsugamushi disease in Shandong region was similar to Kawasaki type in Japan. .