引起儿童腹泻的沙门菌属临床分离株的耐药特点及分子流行病学研究

目的 研究儿童腹泻患者粪便分离的沙门菌属临床分离株的耐药特点及分子流行病学特征.方法 从儿童腹泻患者粪便中分离沙门菌72株,采用血清学凝集试验确定沙门菌血清型;采用K-B纸片扩散法检测抗菌药物的敏感性;采用琼脂稀释法测定头孢噻肟(CTX)和头孢他啶(CAZ)的MIC值;ESBL、ISEcpI和AmpC基因采用PCR法和DNA测序法;采用接合试验测定耐药基因的转移性;PFGE法测定鼠伤寒沙门菌的同源性.结果 从感染性腹泻患儿粪便中共分离出72株沙门菌,其中鼠伤寒沙门菌为主要血清型,占86%(62/72).鼠伤寒沙门菌和汤普森沙门菌通常对临床常用抗菌药物耐药.其中鼠伤寒沙门菌对氨苄西林的耐药率最高(90%,56/62),其次是四环素(81%,50/62),甲氧苄啶/磺胺甲基异(噁)唑(74%,46/62)和氯霉素(66%,41/62).17株(27%,17/62)鼠伤寒沙门菌和2株汤普森沙门菌对CTX耐药.对氨苄西林耐药的49株鼠伤寒沙门菌和2株汤普森沙门菌经PCR扩增后并测序为blaTEM-1b,62株鼠伤寒沙门菌中ESBL基因阳性的有13株(21%,13/62),且主要是blaCTX-M型,其中8株为blaCTX-M-14,3株为blaCTX-M-15,1株为blaCTX-M-55,另外1株为同时blaCTX-M-14和blaCTX-M-55阳性.所有blaCTX-M阳性的菌株均检测出存在上游的插入序列ISEcpl.在1株对头孢西丁耐药的汤普森沙门菌检测出blaDHA-1型AmpC.62株鼠伤寒沙门菌经PFGE分型,发现A型和D型是最主要两个克隆,分别占19%(12/62)和50%(31/62).7株产blaCTX-M型ESBL基因的菌株属于D型.结论 鼠伤寒沙门菌和汤普森沙门菌耐药性严重且blaCTX-M型ESBL基因检出率高;在沙门菌属中发现blaCTX-M-55,在汤普森沙门菌中发现blaDHA-1°克隆性传播是造成鼠伤寒沙门菌流行的主要原因.

Abstract：

Objective To investigate molecular epidemiology and antimicrobial susceptibility of Salmonella spp. isolates recovered from the stool samples of children with diarrhea. Methods Seventy-two isolates of Salmonella spp. were collected from children with diarrhea. The serum type of Salmonella spp.was determined by serology agglutinating method. Antimicrobial susceptibility was determined by K-B disk diffusion method and MICs of cefotaxime and ceftazidime were measured by agar dilution method for Salmonella spp. isolates. PCR and DNA sequencing were used for detecting ESBL, ISEcpl and AmpC genes; The transfer of cefotaxime resistance was determined by conjugation experiments. PFGE was performed for determining the homogeneity of the S. typhimurium isolates. Results A total of 72 isolates of Salmonella spp. were collected, among which S. typhimurium accounted for 86 % (62/72) and was the main serum type. S. typhimurium isolates and S. thompson isolates were often resistant to most of clinically used antimicrobial agents. Resistance of S. thompson isolates to ampicillin was the highest (90%, 56/62),followed by tetracycline (81%, 50/62), trimethoprim/sulfamethoxazole (74%, 46/62) and chloramphenicol (66%, 41/62). Seventeen S. typhimurium isolates (27%, 17/62) and two S. thompson isolates were resistant to cefotaxime. Forty-nine S. typhimurium isolates and two S. thompson isolates were positive for blaTEB-1b and resistant to ampicillin. Thirteen ESBL-producing S. typhimurium isolates (21%, 13/62) were positive for blaCTX-M (eight for blaCTX-M-14, three for blaCTX-M-15, one for blaCTX-M-55, one for both blaCTX-M-14 and blaCTX-M-55). All isolates harboring blaCTX-M genes were positive for upstream insert sequence ISEcpl. blaDHA-1was detected in a cefoxitin-resistant S. thompson isolate. Two main clones (PFGE type A and D) accounting for 19% (12/62) and 50% (31/62) respectively were found among 62 S. typhimurium isolates. Seven CTXM-producing isolates belonged to PFGE type D. Conclusions The multi-resistance to antimicrobial agents and high prevalence of blaCTX-M genes are found among S. typhimurium and S. thompson clinical isolates. blaCTX-M-55 is first found in S. typhimurium isolates and blaDHA-1 is found in S. thompson isolates. Clonal spread is responsible for the dissemination of S. typhimurium isolates.

Molecular epidemiology and antimicrobial susceptibility of Salmonella isolates recovered from children with diarrhea

Objective To investigate molecular epidemiology and antimicrobial susceptibility of Salmonella spp. isolates recovered from the stool samples of children with diarrhea. Methods Seventy-two isolates of Salmonella spp. were collected from children with diarrhea. The serum type of Salmonella spp.was determined by serology agglutinating method. Antimicrobial susceptibility was determined by K-B disk diffusion method and MICs of cefotaxime and ceftazidime were measured by agar dilution method for Salmonella spp. isolates. PCR and DNA sequencing were used for detecting ESBL, ISEcpl and AmpC genes; The transfer of cefotaxime resistance was determined by conjugation experiments. PFGE was performed for determining the homogeneity of the S. typhimurium isolates. Results A total of 72 isolates of Salmonella spp. were collected, among which S. typhimurium accounted for 86 % (62/72) and was the main serum type. S. typhimurium isolates and S. thompson isolates were often resistant to most of clinically used antimicrobial agents. Resistance of S. thompson isolates to ampicillin was the highest (90%, 56/62),followed by tetracycline (81%, 50/62), trimethoprim/sulfamethoxazole (74%, 46/62) and chloramphenicol (66%, 41/62). Seventeen S. typhimurium isolates (27%, 17/62) and two S. thompson isolates were resistant to cefotaxime. Forty-nine S. typhimurium isolates and two S. thompson isolates were positive for blaTEB-1b and resistant to ampicillin. Thirteen ESBL-producing S. typhimurium isolates (21%, 13/62) were positive for blaCTX-M (eight for blaCTX-M-14, three for blaCTX-M-15, one for blaCTX-M-55, one for both blaCTX-M-14 and blaCTX-M-55). All isolates harboring blaCTX-M genes were positive for upstream insert sequence ISEcpl. blaDHA-1was detected in a cefoxitin-resistant S. thompson isolate. Two main clones (PFGE type A and D) accounting for 19% (12/62) and 50% (31/62) respectively were found among 62 S. typhimurium isolates. Seven CTXM-producing isolates belonged to PFGE type D. Conclusions The multi-resistance to antimicrobial agents and high prevalence of blaCTX-M genes are found among S. typhimurium and S. thompson clinical isolates. blaCTX-M-55 is first found in S. typhimurium isolates and blaDHA-1 is found in S. thompson isolates. Clonal spread is responsible for the dissemination of S. typhimurium isolates.