CAJ | 학술논문

目的探讨湖南省CTX-M型超广谱B内酰胺酶(ESBLs)基因型分布情况和变性高效液相色谱(DHPLC)方法检测CTX-M型ESBLs基因型的准确性.方法用多重PCR扩增标准菌株和ESBLs表型阳性的临床菌株blaCTX-M基因,扩增产物经DHPLC分析得到标准菌株和临床菌株色谱峰图,通过比对标准色谱峰图对临床菌株进行基因分型,同时采用单纯随机抽样法选择25株多重PCR扩增阳性菌株进行特异PCR扩增,其产物冉进行基因测序来评估DHPLC法的准确性.结果 142株产ESBLs的肠杆菌科细菌经多重PCR扩增证实109株携带blaCTX-M基因,检出率为76.8%(109/142).109株扩增阳性的菌株经DHPLC分析后检出4种不同的blaCTX-M基因型:33株携带CTX-M-3、19株携带CTX-M-15、5株携带CTX-M-9、52株携带CTX-M-14.25株基因测序结果与DHPLC基因分型结果作比较显示:24株DHPLC的基因分型结果与基因测序结果完全吻合,但有1株DHPLC基因分型为CTX-M-15,而测序为CTX-M-82.结论 DHPLC可对耐药进行基因快速基因分型,具有准确、快速和经济等优点.
목적 탐토호남성CTX-M형초엄보B내선알매(ESBLs)기인형분포정황화변성고효액상색보(DHPLC)방법검측CTX-M형ESBLs기인형적준학성.방법 용다중PCR확증표준균주화ESBLs표형양성적림상균주blaCTX-M기인,확증산물경DHPLC분석득도표준균주화림상균주색보봉도,통과비대표준색보봉도대림상균주진행기인분형,동시채용단순수궤추양법선택25주다중PCR확증양성균주진행특이PCR확증,기산물염진행기인측서래평고DHPLC법적준학성.결과 142주산ESBLs적장간균과세균경다중PCR확증증실109주휴대blaCTX-M기인,검출솔위76.8%(109/142).109주확증양성적균주경DHPLC분석후검출4충불동적blaCTX-M기인형:33주휴대CTX-M-3、19주휴대CTX-M-15、5주휴대CTX-M-9、52주휴대CTX-M-14.25주기인측서결과여DHPLC기인분형결과작비교현시:24주DHPLC적기인분형결과여기인측서결과완전문합,단유1주DHPLC기인분형위CTX-M-15,이측서위CTX-M-82.결론 DHPLC가대내약진행기인쾌속기인분형,구유준학、쾌속화경제등우점.
Objective To investigate the genotype distribution of CTX-M-type extended-spectrum β-1actamase(ESBLs) by denaturing high-performance liquid chromatography(DHPLC) in Hunan Province and the accuracy of DHPLC assay. Methods The blaCTX-M genes of standard strains and clinical ESBLs-producing Enterobacteriaceae were amplified by multiplex PCR followed by DHPLC and genotype determination. 25 isolates randomly selected were sequenced to assess the accuracy of DHPLC method. Results Among 142 ESBLs-producing isolates, 109 isolates carried blaCTX-M gene (76. 8% ). Four different CTX-M genotypes were detected by DHPLC, including CTX-M-3 (33 isolates), CTX-M-15 (19 isolates), CTX-M-14 (52 isolates) and CTX-M-9 (5 isolates). The DHPLC typing of 25 isolates suggested that 24 isolates were verified uniformly by the sequencing, but one CTX-M-15 isolate typed by DHPLC was shown to be CTX-M-82 by sequencing. Conclusion DHPLC is a powerful tool for genotyping of the resistance gene and is worth being applied in the clinical and scientific research with accurate, rapid and economic advantages.