中华微生物学和免疫学杂志
中華微生物學和免疫學雜誌
중화미생물학화면역학잡지
CHINESE JOURNAL OF MICROBIOLOGY AND IMMUNOLOGY
2008年
12期
1109-1115
,共7页
王树坤%姚云波%储从家%山德生%阚飙%刁保卫%吴强%杨汝松%刘红雁%曾丽萍
王樹坤%姚雲波%儲從傢%山德生%闞飆%刁保衛%吳彊%楊汝鬆%劉紅雁%曾麗萍
왕수곤%요운파%저종가%산덕생%감표%조보위%오강%양여송%류홍안%증려평
肠沙门菌副伤寒血清型%抗微生物药物敏感性%脉冲场凝胶电泳%克隆扩散%遗传多样性
腸沙門菌副傷寒血清型%抗微生物藥物敏感性%脈遲場凝膠電泳%剋隆擴散%遺傳多樣性
장사문균부상한혈청형%항미생물약물민감성%맥충장응효전영%극륭확산%유전다양성
Salmonella enterica serotype Paratyphi%Antimicrobial susceptibility%Pulsed-field gel electrophoresis%Clonal expansion%Genetic diversity
目的 认识肠沙门菌甲型副伤寒血清型(SPA)的克隆扩散和遗传多样性,建立并确定病原菌流行克隆的分型方法.方法 采用有对照的K-B纸片扩散技术对分离的3980株SPA进行抗微生物药物敏感性试验;经PCR扩增和基因测序检测15个萘啶酸抗性菌株喹诺酮抗性决定区的gyrA、gyrB、syrC和syrE基因;采用Spe Ⅰ和Xba Ⅰ消化染色体DNA脉冲场凝胶电泳(PFGE)对来自7个县的121个分离株进行分型和聚类分析.结果 萘啶酸敏感菌株在1999年占有优势,但2000年以后被萘啶酸抗性菌株替代;15个萘啶酸抗性菌株的PCR扩增和基因测序显示抗性机制是由gyrA基因的单点突变引起;121个菌株spe Ⅰ和Xba Ⅰ消化产物分别得出以Spe Ⅰ 01、spe Ⅰ 02或Xba Ⅰ 01型占优势的5种和4种PFGE型,Spe Ⅰ 01和Spe Ⅰ 02分别占37.2%和57.9%,或Xhn Ⅰ 01占95.0%.结论 在研究期间SPA分离株萘啶酸抗性率上升;PFGE型的SpeⅠ01和SpeⅠ 02或XbaⅠ01是玉溪流行的主要克隆;采用Spe Ⅰ和Xba Ⅰ的PFGE是鉴别SPA的一项有用技术.
目的 認識腸沙門菌甲型副傷寒血清型(SPA)的剋隆擴散和遺傳多樣性,建立併確定病原菌流行剋隆的分型方法.方法 採用有對照的K-B紙片擴散技術對分離的3980株SPA進行抗微生物藥物敏感性試驗;經PCR擴增和基因測序檢測15箇萘啶痠抗性菌株喹諾酮抗性決定區的gyrA、gyrB、syrC和syrE基因;採用Spe Ⅰ和Xba Ⅰ消化染色體DNA脈遲場凝膠電泳(PFGE)對來自7箇縣的121箇分離株進行分型和聚類分析.結果 萘啶痠敏感菌株在1999年佔有優勢,但2000年以後被萘啶痠抗性菌株替代;15箇萘啶痠抗性菌株的PCR擴增和基因測序顯示抗性機製是由gyrA基因的單點突變引起;121箇菌株spe Ⅰ和Xba Ⅰ消化產物分彆得齣以Spe Ⅰ 01、spe Ⅰ 02或Xba Ⅰ 01型佔優勢的5種和4種PFGE型,Spe Ⅰ 01和Spe Ⅰ 02分彆佔37.2%和57.9%,或Xhn Ⅰ 01佔95.0%.結論 在研究期間SPA分離株萘啶痠抗性率上升;PFGE型的SpeⅠ01和SpeⅠ 02或XbaⅠ01是玉溪流行的主要剋隆;採用Spe Ⅰ和Xba Ⅰ的PFGE是鑒彆SPA的一項有用技術.
목적 인식장사문균갑형부상한혈청형(SPA)적극륭확산화유전다양성,건립병학정병원균류행극륭적분형방법.방법 채용유대조적K-B지편확산기술대분리적3980주SPA진행항미생물약물민감성시험;경PCR확증화기인측서검측15개내정산항성균주규낙동항성결정구적gyrA、gyrB、syrC화syrE기인;채용Spe Ⅰ화Xba Ⅰ소화염색체DNA맥충장응효전영(PFGE)대래자7개현적121개분리주진행분형화취류분석.결과 내정산민감균주재1999년점유우세,단2000년이후피내정산항성균주체대;15개내정산항성균주적PCR확증화기인측서현시항성궤제시유gyrA기인적단점돌변인기;121개균주spe Ⅰ화Xba Ⅰ소화산물분별득출이Spe Ⅰ 01、spe Ⅰ 02혹Xba Ⅰ 01형점우세적5충화4충PFGE형,Spe Ⅰ 01화Spe Ⅰ 02분별점37.2%화57.9%,혹Xhn Ⅰ 01점95.0%.결론 재연구기간SPA분리주내정산항성솔상승;PFGE형적SpeⅠ01화SpeⅠ 02혹XbaⅠ01시옥계류행적주요극륭;채용Spe Ⅰ화Xba Ⅰ적PFGE시감별SPA적일항유용기술.
Objective To understand the elonal expansion and genetic diversity of Salmonella en-terica semtype Paratyphi A (SPA) and to construct a typing method to determine the epidemic clones of the isolates. Methods Antimicrobial susceptibility testing was performed with 3980 SPA isolates by the cen-trolled Kirby-Bauer disc diffusion technique on Muller-Hinton agar plates. A total of 15 SPA with nalidixie acid resistance for mutations in gyrA, gyrB, gyrC and gyrE genes within the quinolone-resistant determina-tion region (QRDR) were examined. Subtyping of 121 isolates of SPA from seven counties in Yuxi were studied using pulsed-field gel eleetrophoresis (PFGE) analysis following digestion of chromosomal DNA with restriction endanucleases Spe Ⅰ and Xba Ⅰ. PFGE patterns were analyzed by duster analysis. Results The nalidixic acid-susceptible isolates predominated in 1999 but was replaced by nalidixic acid -resistant (NAR) isolates after 2000. Amplification by PCR and sequencing of the genes with subsets of 15 NAR strains re-vealed that the resistance mechanisms had resulted from single point mutations in the gyrA gene. Spe Ⅰ and Xba Ⅰ digestion of 121 isolates gave five and four different PFGE patterns with the predominance of the Spe Ⅰ 01 and Spe Ⅰ 02 (or the Xba Ⅰ 01) epidemic patterns, respectively. Spe Ⅰ 01 and Spe Ⅰ 02 consisted of 37.2% and 57.9% of isolates, respectively, or Xba Ⅰ 01 consisted of 95.0% of isolates. Conclusion The incidence of resistance to nalidixic acid of the isolates increased during the study period. PFGE patterns Spe Ⅰ 01 and Spe Ⅰ 02 (or Xba Ⅰ 01), the main clones of the epidemics, are highly prevalent in Yuxi. PFGE with Spe Ⅰ and Xba Ⅰ is a useful technique to differentiate SPA.
