大肠埃希菌ESBLs基因型与生物被膜调控基因pgaABCD对其耐药性的相关性分析  

Correlation analysis of extended spectrum beta-lactamases(ESBLs)gene and biofilm regulatory gene pgaABCD on drug resistance of Escherichia coli(E.coli)

作　　者：马尔马尔·托乎塔尔别克 热汗古力·玉苏甫 熊文娟 湛文博 吴丽萍[1] Maermaer Tuohutaerbeike;Rehanguli Yusufu;XIONG Wenjuan;ZHAN Wenbo;WU Liping(Department of the Laboratory,the Fifth Affiliated Hospital of Xinjiang Medical University,Urumqi 830011,China)

机构地区：[1]新疆医科大学第五附属医院检验科,乌鲁木齐830011

出　　处：《新疆医科大学学报》2025年第2期141-148,共8页Journal of Xinjiang Medical University

基　　金：新疆少数民族科技人才特殊培养计划科研项目(2021D03020)。

摘　　要：目的探究产超广谱β-内酰胺酶(Extended spectrum beta-lactamases,ESBLs)基因与生物被膜调控基因pgaABCD表达之间的相关性以及二者对产ESBLs大肠埃希菌抗生素耐药性的影响。方法收集2021年1-12月本院临床分离的大肠埃希菌,通过双纸片协同试验筛选出产ESBLs菌株66株,采用标准Kirby-Bauer圆盘扩散法进行药敏试验,通过实时荧光PCR鉴定ESBLs基因bla_(CTX-M-1)、bla_(CTX-M-14)、bla_(TEM-1)、bla_(SHV-12)和生物被膜调控基因pgaA、pgaB、pgaC、pgaD的分布情况,采用结晶紫染色法评价生物被膜形成能力,对比不同生物被膜形成能力组间生物被膜调控基因表达差异,对ESBLs基因检出率、pgaABCD基因检出率与生物被膜形成能力之间进行相关性分析。结果66株产ESBLs大肠埃希菌的耐药模式以多重耐药为主:75.8%的分离株对3种及以上抗生素耐药,耐药率前三的抗生素是氨苄西林(100.0%)、头孢唑林(100.0%)和头孢曲松(97.0%),其次是左氧氟沙星和环丙沙星(耐药率均为87.9%);所有菌株均能够形成生物被膜,分别有40株(60.6%)、22株(33.3%)和4株(6.1%)菌株表现出强、中和弱水平生物被膜形成能力;检出最多的ESBLs基因型是bla_(CTX-M-1)(57.6%),其次是bla_(TEM-1)(39.4%),有30株菌株同时携带2种及以上ESBLs基因,所有受试菌均检出pgaA基因,60株菌检出pgaC基因,36株菌同时携带pgaA、pgaB、pgaC和pgaD 4种基因,其中pgaC基因的检出率分别与生物被膜形成能力(Phi=0.367,P=0.012)及bla_(CTX-M-1)基因检出率(Phi=0.368,P=0.003)呈显著正相关。生物被膜形成能力与抗生素耐药性之间虽未发现显著相关性(P>0.05),但表现出生物被膜形成能力越强,菌株携带的ESBLs基因种类越多,耐药特征也愈加复杂的特点。结论bla_(CTX-M-1)可能通过上调生物被膜调控基因pgaC的表达增强生物被膜形成能力,从而促进耐药基因传播。Objective To explore the correlation between the expression level of extended spectrum betalactamases(ESBLs)genes and the biofilm regulatory genes pgaABCD and their effcts on the antibiotic resistance of ESBLs-producing Escherichia coli(E.coli).Methods A total of 66 ESBLs-producing strains were screened in a double-disc synergy test on clinical isolates of E.coli from January to December 2021 in the hospital,and drug sensitivity tests were performed using the standard Kirby-Bauer disc diffusion method,the distribution of ESBL genotypes bla_(CTX-M-1),bla_(CTX-M-14),bla_(TEM-1),bla_(SHV-12)and biofilm genes pgaA,pgaB,pgaC,pgaD were identified by real-time fluorescence PCR.The biofilm-forming ability was assessed by crystalline violet staining.The research compared the differences in the expression of biofilm genes among the groups with different biofilm-forming abilities and analyzed the correlation between ESBLs gene detection rate,pgaABCD gene detection rate and biofilm formation ability.Results The resistance pattern of the 66 ESBLs-producing E.coli strains was dominated by multi-resistance:75.8%of the isolates were resistant to 3 or more antibiotics,and the top 3 antibiotics were ampicillin(100.0%),ceftaroline(100.0%)and ceftriaxone(97.0%),followed by levofloxacin and ciprofloxacin(both with a rate of resistance of 87.9%).All 66 isolates were able to form biofilms of which 40(60.6%),22(33.3%)and 4(6.1%)isolates showed strong,moderate and weak levels of biofilm formation,respectively.The most prevalent ESBLs genotype in this study was bla_(CTX-M-1)(57.6%),followed by bla_(TEM-1)(39.4%),there were also 30 strains carried 2 or more ESBLs genes.PgaA gene was detected in all the subjects,pgaC gene was detected in 60 strains,and 36 strains carried pgaA,pgaB,pgaC and pgaD genes,in which the detection rate of pgaC gene showed a significant positive correlation with the biofilm formation ability(Phi=0.367,P=0.012)and bla_(CTX-M-1)gene detection rate(Phi=0.368,P=0.003),respectively.Although no significant correlation was

关 键 词：大肠埃希菌 产超广谱β-内酰胺酶(ESBLs) 生物被膜 pga基因 耐药性 

分 类 号：R446.5[医药卫生—诊断学]