复发性尿路感染患者致病性大肠埃希菌分子分型、毒力基因及药物敏感性分析

doi:10.3969/j.issn.1673-8640.2022.02.009

摘要/Abstract

摘要：

目的分析复发性尿路感染（rUTI）患者致病性大肠埃希菌分子分型、毒力基因和耐药性。方法收集分离自上海市第六人民医院60例rUTI患者的大肠埃希菌菌株,采用脉冲场凝胶电泳（PFGE）、多位点序列分型（MLST）和种系发育分型方法进行克隆分型,采用聚合酶链反应（PCR）和测序法分析其常见毒力基因（feoB、chuA、iutA、iroN、ireA、fimH、foc、sfa、aer和cnf）,并分析大肠埃希菌对13种常用抗菌药物的耐药性。结果根据PFGE结果,将60例rUTI患者分为复发组和再感染组,纳入分离自复发组的30株和再感染组的60株大肠埃希菌。纳入菌株的主要MLST型别为ST131型和ST405型,主要种系发育分型为B2型和D型。在10种毒力基因中,feoB所占比例最高（98.8%）,其次是fimH（93.3%）。复发组iutA携带率高于再感染组（P=0.020）。2个组临床分离菌株抗菌药物耐药性模式无明显差异（P>0.05）。结论 ST131和ST405是上海市第六人民医院rUTI患者致病性大肠埃希菌的主要MLST型别,常见毒力基因为feoB和fimH,iutA在复发患者中携带率较高,可将其作为降低尿路感染持留的治疗靶点。

关键词: 尿路致病性大肠埃希菌, 复发性尿路感染, 脉冲场凝胶电泳, 毒力基因

Abstract:

Objective To investigate molecular typing,virulence genes and antibiotic susceptibility of uropathogenic Escherichia coli in patients with recurrent urinary tract infection（rUTI）. Methods Totally,60 patients with rUTI were enrolled,and the isolates of Escherichia coli were isolated. Pulsed-field gel electrophoresis（PFGE）,multi-locus sequence typing（MLST）and phylogenetic groups were performed. Polymerase chain reaction（PCR） and sequencing were used to analyze common Escherichia coli virulence genes（feoB,chuA,iutA,iroN,ireA,fimH,foc,sfa,aer and cnf）. The antibiotic susceptibilities of Escherichia coli to 13 common antimicrobial agents were analyzed. Results The research subjects were classified into relapse and reinfection groups by PFGE. A total of 30 isolates from relapse group and 60 isolates from reinfection group were collected for further analysis. MLST indicated that ST131 was the predominant type followed by ST405. The major phylogenetic groups were B2 and D. Among the 10 virulence genes,feoB（98.8%） was the most common virulence gene followed by fimH（93.3%）. Except for iutA（P=0.020）,no statistical significance of the other virulence genes was found between the 2 groups（P>0.05）. Conclusions The predominant types of uropathogenic Escherichia coli are ST405 and ST131 in Shanghai Sixth People's Hospital. Two virulence genes,feoB and fimH,are highly prevalent. The carry rate of iutA is high in relapse group,which can be as the treatment target for rUTI.

Key words: Uropathogenic Escherichia coli, Recurrent urinary tract infection, Pulsed-field gel electrophoresis, Virulence gene

中图分类号:

R446.1

潘允琪, 李运改, 王坚镪, 吴琼, 汤瑾. 复发性尿路感染患者致病性大肠埃希菌分子分型、毒力基因及药物敏感性分析[J]. 检验医学, 2022, 37(2): 141-145.

PAN Yunqi, LI Yungai, WANG Jianqiang, WU Qiong, TANG Jin. Analysis of molecular typing,virulence genes and antibiotic susceptibility of uropathogenic Escherichia coli in patients with recurrent urinary tract infection[J]. Laboratory Medicine, 2022, 37(2): 141-145.

图/表 4

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基因或DNA片段	碱基长度/bp	引物序列（5'~ 3'）	退火温度/ ℃
chuA	1 500	F:GCTGTTTCTGCCACCTTG	55
		R:CCTGAGTTTCGTTAGTTTCC
yjaA	211	F:TGAAGTGTCAGGAGACGCTG	55
		R:ATGGAGAATGCGTTCCTCAAC
TSPE4.C2	152	F:GAGTAATGTCGGGGCATTCA	53
		R:CGCGCCAACAAAGTATTACG

基因或DNA片段	碱基长度/bp	引物序列（5'~ 3'）	退火温度/ ℃
chuA	1 500	F:GCTGTTTCTGCCACCTTG	55
		R:CCTGAGTTTCGTTAGTTTCC
yjaA	211	F:TGAAGTGTCAGGAGACGCTG	55
		R:ATGGAGAATGCGTTCCTCAAC
TSPE4.C2	152	F:GAGTAATGTCGGGGCATTCA	53
		R:CGCGCCAACAAAGTATTACG

组别	菌株数/株	A		B1		B2		D		合计
组别	菌株数/株	菌株数/株	百分比/%	菌株数/株	百分比/%	菌株数/株	百分比/%	菌株数/株	百分比/%	菌株数/株	百分比/%
再感染组	60	2	3.3	15	25.0	12	20.0	31	51.7	60	100.0
复发组	30	1	3.3	4	13.3	6	20.0	19	63.3	30	100.0

组别	菌株数/株	A		B1		B2		D		合计
组别	菌株数/株	菌株数/株	百分比/%	菌株数/株	百分比/%	菌株数/株	百分比/%	菌株数/株	百分比/%	菌株数/株	百分比/%
再感染组	60	2	3.3	15	25.0	12	20.0	31	51.7	60	100.0
复发组	30	1	3.3	4	13.3	6	20.0	19	63.3	30	100.0

组别	黏附基因			铁获取基因						毒力基因 cnf
组别	fimH	sfa	foc	foeB	chuA	ireA	iutA	iroN	aer	毒力基因 cnf
再感染组	27（90.0）	1 （3.3）	6 （20.0）	29（96.7）	25（83.3）	3 （10.0）	23（76.7）	19（63.3）	20（66.7）	0 （0）
复发组	57（95.0）	2 （3.3）	7 （11.7）	60（100.0）	44（73.3）	10（16.7）	55（91.7）	36（60.0）	40（66.7）	5 （8.3）
P值	0.396	1.000	0.289	0.333	0.290	0.396	0.020	0.760	1.000	0.165