宁波市耐三代头孢菌素志贺菌携带β-内酰胺酶与基因分型

Gene typing of ESBLs-producing third-generation cephalosporins-resistant Shigellain Ningbo,China

目的了解本市耐三代头孢菌素志贺菌携带ESBLs及其基因型,为疾病防控提供依据。方法采用直接和增菌培养法,分离患者标本中志贺菌;药敏采用K-B法,ESBLs志贺菌表型确证采用纸片法;CTX-M、OXA、TEM和SHV耐药基因采用PCR法;耐药基因分型采用核苷酸序列法,用BLAST分析比较确定基因型别。结果药敏筛检出69株耐三代头孢菌素志贺菌,占ESBLs志贺菌的74.19%。检出CTX-M(CTX-M-1群和CTX-M-9群)、OXA和TEM耐药基因,检出率分别为79.71%、79.01%和26.09%,未检出CTX-M-2群和SHV耐药基因。DNA序列比对CTX-M-1群以CTX-M-15型为主,还检出7个其它型;CTX-M-9群则以CTX-M-14型多,其它型检出6个;49株OXA和18株TEM耐药基因测序后均为1型(OXA-1型和TEM-1型)。携带2种以上耐药基因的志贺菌21株,占30.43%。结论本市志贺菌对头孢曲松等三代头孢菌素耐药率较高,检出的ESBLs酶型种类多,CTX-M(CTX-M-1群的CTX-M-15型和CTX-M-9群的CTX-M-14型)是主流酶型,且可同时携带多种耐药基因,给疾病防控带来困难。OXA-1型的高携带率,提示我们应加强分析。B群志贺菌无论是表型的耐药性,还是耐药基因检出率均高于D群志贺菌,该发现有助志贺菌的扩散与流行研究。

We investigated the third-generation cephalosporins-resistant Shigellaand its genotype in Ningbo,China,providing a basis for disease prevention and control.Pathogenic bacteria were analyzed by direct isolation combined with enrichment culture isolation.Antimicrobial susceptibility was determined by K-B disk diffusion method and PCR was used for detecting multidrug resistance genes like CTX-M,OXA,TEMand SHV.BLAST analysis was used to determine the genotype.Results showed that 69 strains of third-generation cephalosporins-resistant Shigella were detected by drug sensitivity screening,accounting for 74.19% of ESBLs Shigella.Drug resistance gene CTX-M（CTM-M-1and CTM-M-9）,OXAand TEM were detected.The detection rate were 79.71%,79.01% and 26.09% respectively.With no CTX-M-2and SHV,DNA sequence alignment showed CTX-M-1group were mainly of CTX-M-15 type besides seven other types;CTX-M-9group were mainly of CTX-M-14 type besides six other types;49strains of OXAand 18 strains of TEM were sequenced to be type 1（OXA-1and TEM-1type）.The 21 Shigellastrains carrying more than two drug resistance genes accounts for 30.43%.Shigellain Ningbo has high third-generation cephalosporins-resistance rate and many kinds of ESBLs enzymes were detected.The mainstream enzyme type was CTX-M,meanwhile they also carried a variety of drug resistance genes,which could bring difficulties to disease prevention and control.The high carrying rate of OXA-1type suggests that we should pay more attention.The detection rate of group B was higher than that of group D,including not only the phenotype resistance but also the drug-resistance genes;these findings will be useful in the study of the drug resistance prevalence of Shigella.