摘要
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 。
Aim To determine genotypes of Orientia tsutsugamushi isolated from the Nanpeng islands.Methods The 56 kD protein coding regions of Orientia tsutsugamudshi isolated 8 strains from the Nanpeng islands were amplified by nested polymerase chain reaction (NPCR) and characterized by RFLP.The purified positive products of 4 strains were cloned into pGEM-T vector and sequenced.The DNA sequences were compared with various sequences on the internet for sequence homology analysis.Results 500 bp DNA fragments encoding 56-kD protein were amplified in 7 of 8 samples.Results of RFLP showed that three strains existed.Among 7 clones from 4 samples,two (DY1,DY21) were almost identical to Yonchon strain(with nucleotide sequence homology of 99%),another three (DY22,NY12,NS2) were similar to Karp strain(92%),and the other two(NY11,NS1) were homologous to Kato strain(98%).Conclusion There existed three genotypes of Orientia tsutsugamushi which were Karp,Kato,and Yonchon on the Nanpeng islands.
出处
《中国人兽共患病杂志》
CSCD
北大核心
2001年第3期43-45,共3页
Chinese Journal of Zoonoses
基金
总后勤部卫生部九.五重点课题! (编号 97L0 5 9)
