应用PCR/RFLP对山东地区恙虫病东方体的基因型分析  

Analysis of the gene type of Orientia tsutsugamushi isolated from Shandong by PCR/RFLP

作　　者：刘运喜[1] 赵仲堂[2] 高媛 张景兰 杨占清 步秀萍 苏静静 

机构地区：[1]济南军区军事医学研究所流行病研究室,济南250014 [2]山东大学公共卫生学院流行病学与卫生统计学研究所

出　　处：《中国人兽共患病杂志》2004年第5期425-428,共4页Chinese Journal of Zoonoses

基　　金：国家自然科学基金 (No 3 0 3 712 3 7);解放军总后勤部卫生部全军医药卫生科研基金 (No 0 1Q0 16);山东省科学技术发展计划(No 0 2 2 13 0 111)资助项目

摘　　要：目的　鉴定山东地区恙虫病东方体的基因型。方法　采用聚合酶链反应 /限制性片段长度多态性分析 (PCR/RFLP)技术对恙虫病东方体山东分离株、小盾纤恙螨匀浆及恙虫病病人全血中提取的DNASta5 6基因进行分析研究 ,并与NestedPCR基因分型的结果进行比较。结果　山东地区 35份标本 ( 2 3株恙虫病东方体山东分离株、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 )提取的DNA群引物扩增产物经HinfⅠ、HhaⅠ、SnaBⅠ酶切后呈现 2种不同的RFLP图谱 ,经与已知序列的Ot内切酶图谱相比 :33份Ot内切酶图谱与文献报告的日本地方株—Kawasaki株具有相似的酶切图谱 ,但缺乏HhaⅠ的酶切位点 ;另 2份与Karp参考株具有相同的酶切图谱。该结果与NestedPCR基因分型的结果相一致。结论　山东地区恙虫病东方体流行株主要基因型与日本地方株Kawasaki型类似 ,但与日本地方株存在遗传差异 ;同时还有Karp型Ot存在。采用PCR/RFLP方法可以准确对Ot-Sta5 6基因分型。Aim In order to identify the gene type of Orientia tsutsugamushi(Ot) isolated from Shandong province Method RFLP analysis was used to analyze restriction profiles of the amplified products of Ot Sta56 gene of the isolates by group primers,and the results were compared with those acquired by Nested PCR Results of the 35 samples (23strains Ot isolated from scrub typhus patients(8 strains),chigger mites [4strains from Leptotrombidium (L ) scutellare;3 strains from L palpale;and 2strains from L linhuaikongense and Walchia (W ) pacifica respectively],and wild rodents (4 strains from Apodemus (A ) agrarius) in Shandong province,10 samples of scrub patients blood,and 2 pools of homogenized L scutellare) 33 had the similar restriction maps with Japan Kawasaki ,but they had no restriction site of HhaⅠ,and thus had some differences in gene sequence in comparison;2 had the similar restriction maps with Karp This results were identical to that acquired by Nested PCR Conclusion The genotypes of epidemic Ot in Shandong province were similar to Japan Kawasaki,but had some differences in gene sequence,in addition,Karp also existed among rodents and chigger mites in this region PCR/RFLP could be used to accurately analyze the gene type of Ot

关 键 词：聚合酶链反应/限制性片段长度多态性分析(PCR/RFLP) 恙虫病东方体 基因型 

分 类 号：R376.2[医药卫生—病原生物学]