CAJ | 학술논문

目的：鉴定莫西沙星耐药的艰难梭菌临床菌株的基因型和毒素相关基因的携带情况。方法22株莫西沙星耐药的艰难梭菌临床菌株收集自澳大利亚悉尼地区，利用基于荧光毛细管电泳的聚合酶链反应（PCR）‐核糖体分型技术对其进行基因分型；利用PCR方法检测其毒素A、B编码基因 tcdA、tcdB和二元毒素编码基因cdtA 、cdtB；利用PCR‐基因测序方法检测毒素调节基因tcdC的基因序列，通过与参考菌株VPI 10463（Genbank收录号：X92982）比对了解其基因突变情况，通过与Genbank中的序列比对确定其序列型。结果21株菌株为高毒力核糖体型027（RT027），另1株为RT078；22株菌株毒力编码基因均为 tcdA＋tcdB＋ cdtA＋ cdtB＋；21株RT027 tcdC序列型为sc1，另1株RT078序列型为WA39，与参考株VPI 10463比较，RT027菌株tc‐dC序列均出现连续18个核苷酸（nt330‐347）的缺失和第117位单核苷酸的缺失，RT078菌株则出现连续39个核苷酸（nt341‐379）的缺失和第184位（C＞ T ）的突变。结论高毒力 RT027是最常见的莫西沙星耐药的艰难梭菌基因型；高毒力 RT027和RT078均携带毒素A、B和二元毒素编码基因，且毒素调节基因 tcdC均存在基因突变。
목적：감정막서사성내약적간난사균림상균주적기인형화독소상관기인적휴대정황。방법22주막서사성내약적간난사균림상균주수집자오대리아실니지구，이용기우형광모세관전영적취합매련반응（PCR）‐핵당체분형기술대기진행기인분형；이용PCR방법검측기독소A、B편마기인 tcdA、tcdB화이원독소편마기인cdtA 、cdtB；이용PCR‐기인측서방법검측독소조절기인tcdC적기인서렬，통과여삼고균주VPI 10463（Genbank수록호：X92982）비대료해기기인돌변정황，통과여Genbank중적서렬비대학정기서렬형。결과21주균주위고독력핵당체형027（RT027），령1주위RT078；22주균주독력편마기인균위 tcdA＋tcdB＋ cdtA＋ cdtB＋；21주RT027 tcdC서렬형위sc1，령1주RT078서렬형위WA39，여삼고주VPI 10463비교，RT027균주tc‐dC서렬균출현련속18개핵감산（nt330‐347）적결실화제117위단핵감산적결실，RT078균주칙출현련속39개핵감산（nt341‐379）적결실화제184위（C＞ T ）적돌변。결론고독력 RT027시최상견적막서사성내약적간난사균기인형；고독력 RT027화RT078균휴대독소A、B화이원독소편마기인，차독소조절기인 tcdC균존재기인돌변。
Objective To investigate the genotype and variance of toxin associated genes of moxifloxacin‐resistant Clostridium difficile clinical isolates in Sydney .Methods Twenty‐two moxifloxacin‐resistant Clostridium difficile clinical isolates were collected from Sydney ,which were genotyped by using sequencer capillary gel electrophoresis based PCR‐ribotyping ,and toxin A and B cod‐ing gene tcdA and tcdB ,and binary toxin coding gene cdtA and cdtB were detected by using PCR method .Toxin regulator gene tc‐dC was analyzed by using PCR‐sequencing ,and was aligned with reference sequence of VPI 10463 (Genbank accession number :X92982) ,and the tcdC sequence types of all 22 isolates were identified by using blast tool in NCBI .Results Twenty‐one isolates were genotyped as hypervirulent PCR‐ribotypes 027 (RT027) ,and one isolate as RT078 ;all 22 isolates contained tcdA and tcdB for toxin A and B and cdtA and cdtB for binary toxin (tcdA+ tcdB+ cdtA+ cdtB+ ) .The tcdC sequence types of the 21 RT027 i‐solates belong to sc1 ,and that of the one RT078 isolate belongs to WA39 .Compared with tcdC reference sequence of VPI 10463 ,a consecutive 18 bp deletion (nt341 to 379) and one nucleotide deletion at position 117 were found in the 21 RT027 isolates ,and a consecutive 39 bp deletion (nt330 to 368) and one nucleotide mutation at position 184(C> T) were found in the one RT078 isolate . Conclusion Clostridium difficile hypervirulent RT027 was the common moxifloxacin resistant genotype ;Clostridium difficile hy‐pervirulent RT027 and RT078 clinical isolates contained genes for toxin A and B and binary toxin ,and contained gene sequence mu‐tation in toxin regulator gene tcdC .