CAJ | 학술논문

目的：探索非典型O1群埃尔托型霍乱弧菌在福建省1962-2005年霍乱菌株中的存在及其意义。方法选择1962-2005年代表性霍乱菌株69株（稻叶21、小川48），运用PCR技术进行ctxB、tcpA、rstR、hlyA 各毒力因子的古典型（Cl）和埃尔托型（El）基因的扩增；同时对部分菌株的ctxB和rstR基因的序列进行测定和比对；分析福建省O1群埃尔托型霍乱弧菌（EVC）中非典型菌株出现的时间和存在意义。结果1962年福建省分离的O1群 EVC菌株中携带有 ctxB-Cl、tcpA-Cl、hlyA-Cl基因；rstR-Cl基因仅在1994-2000年的霍乱菌株中检出。序列比对结果发现ctxB基因出现新的204位碱基突变位点（T→G）；同时报道两种新的ctxB基因型别（10和11），且发现存在于同一个菌株中；ctxB基因型11的115位碱基为C ，表现为古典型 ctxB位点特征，203位碱基为T ，表现为埃尔托型ctxB位点特征。结论福建省在1962年的O1群EVC菌株中就出现杂合有 ctxB-Cl、tcpA-Cl、hlyA-Cl基因的非典型EVC菌株，是迄今为止报道出现非典型EVC最早的年代；1994年以后出现整合有 rstR-Cl基因的非典型EVC菌株。另外，在福建省O1群EVC中存在国内外未曾报道的新的 ctxB碱基突变位点和ctxB 基因型别；同时在同一菌株中存在不同的ctxB基因型别杂合形式；ctxB序列也存在不同生物型别的特异位点组合形式。福建省O1群EVC表现为独特的地方性分子特征。
목적：탐색비전형O1군애이탁형곽란호균재복건성1962-2005년곽란균주중적존재급기의의。방법선택1962-2005년대표성곽란균주69주（도협21、소천48），운용PCR기술진행ctxB、tcpA、rstR、hlyA 각독력인자적고전형（Cl）화애이탁형（El）기인적확증；동시대부분균주적ctxB화rstR기인적서렬진행측정화비대；분석복건성O1군애이탁형곽란호균（EVC）중비전형균주출현적시간화존재의의。결과1962년복건성분리적O1군 EVC균주중휴대유 ctxB-Cl、tcpA-Cl、hlyA-Cl기인；rstR-Cl기인부재1994-2000년적곽란균주중검출。서렬비대결과발현ctxB기인출현신적204위감기돌변위점（T→G）；동시보도량충신적ctxB기인형별（10화11），차발현존재우동일개균주중；ctxB기인형11적115위감기위C ，표현위고전형 ctxB위점특정，203위감기위T ，표현위애이탁형ctxB위점특정。결론복건성재1962년적O1군EVC균주중취출현잡합유 ctxB-Cl、tcpA-Cl、hlyA-Cl기인적비전형EVC균주，시흘금위지보도출현비전형EVC최조적년대；1994년이후출현정합유 rstR-Cl기인적비전형EVC균주。령외，재복건성O1군EVC중존재국내외미증보도적신적 ctxB감기돌변위점화ctxB 기인형별；동시재동일균주중존재불동적ctxB기인형별잡합형식；ctxB서렬야존재불동생물형별적특이위점조합형식。복건성O1군EVC표현위독특적지방성분자특정。
The emergence of atypical El Tor strains from V .cholerae in South Asia and Africa has been attributed to several outbreaks in recent decades ,however ,backgrounds of such strains in China remain exclusive .In this study ,PCR am-plification of both El Tor and classical alleles for ctxB ,tcpA ,rstR and hlyA genes was attempted in sixty-nine El Tor isolates from Fujian between 1962 to 2005 ,in addition ,some amplicons were sequence-analyzed .Thus ,the time point of atypical EVC strains in Fujian was determined ,genetic diversities of such strains were investigated .It was revealed that ctxB-Cl ,tcpA-Cl and hlyA genes were detected in O1 serogroup EVC isolates from Fujian since 1962 .Although rstR-Cl gene was solely detected in isolates between 1994 to 2000 .It was indicated by sequence analysis that atypical EVC strains from Fujian possessed a novel T→G mutation at residual 204 of the ctxB gene .Remarkably ,two novel ctxB genotypes (ctxB-10 and ctxB-11) were identified in one strain .The residual 115-C of ctxB in ctxB-11 showed characteristics of ctxB-Cl ,however ,its residual 203-T demonstra-ted characteristics of ctxB-El .This observation implied that it was common in O1 serogroup EVC strains from Fujian hybrid-ized with classical alleles since 1962 ,which would be the earliest time-point for the emergence of atypical El Tor strains hitherto in literature .Emergence of atypical El Tor strains harboring rstR-Cl in Fujian occurred since 1994 .Meanwhile ,novel mutation sites and ctxB genotypes were observed in Fujian isolates ,including diverse combination of ctxB genotypes in one strain and combination of biotype-specific sites in ctxB sequences .In summary ,molecular characterization of O 1 serogroup EVC strains from Fujian was unique and geography-associated .