200株血流感染大肠埃希菌耐药性分析

doi:10.3969/j.issn.1673-8640.2020.12.014

摘要/Abstract

摘要：

目的了解安宁地区产超广谱β-内酰胺酶（ESBL）大肠埃希菌基因分型及其与整合子的关系,掌握大肠埃希菌耐药特点及流行趋势,为临床治疗提供参考。方法收集住院患者大肠埃希菌临床分离株200株,进行菌种鉴定和体外药物敏感性试验。采用纸片扩散法检测ESBL菌株,并通过双纸片法进行确认。采用聚合酶链反应（PCR）检测ESBL基因型别,同时扩增整合子5、3及保守区Ⅰ类、Ⅱ类、Ⅲ类整合酶基因,进行整合子测序分类。结果 200株大肠埃希菌中,有118株（59.00%）检出ESBL阳性,88.98%（105/118）扩增到ESBL基因,9.30%（11/118）株同时检出2种ESBL基因。118株ESBL阳性菌株中,CTX-M型的阳性检出率最高（52.54%）。不同基因型大肠埃希菌中,仅检出Ⅰ类整合子,其中CTX-M-1型携带Ⅰ类整合子的菌株所占比例最高（72.41%）,SHV型携带Ⅰ类整合子的菌株所占比例最低（50.00%）。大肠埃希菌对氨苄西林-舒巴坦的耐药率最高（83.23%）,对亚胺培南的敏感率最高（97.42%）。结论安宁地区大肠埃希菌产ESBL率较高,基因型主要以CXT-M为主,携带多种ESBL基因的菌株主要以携带Ⅰ类整合子为主。

关键词: 大肠埃希菌, 超广谱β-内酰胺酶, 基因分型, 整合子

Abstract:

Objective To understand the relationship between extended-spectrum lactamases（ESBL） genotyping of Escherichia coli and integron in Anning,to evaluate the characteristics and prevalence of Escherichia coli drug resistance,and to provide a reference for the treatment and control of Escherichia coli producing ESBL. Methods A total of 200 isolates of Escherichia coli were collected. Isolate identification was performed,and drug susceptibility test was performed in vitro. ESBL isolates were determined by disc diffusion method and confirmed by double disk method. ESBL genotyping was determined by polymerase chain reaction（PCR）. Integron 5 and 3 and conservative district of Ⅰ,Ⅱ and Ⅲ integrase genes were amplified,and integrates were sequenced and classified. Results Among the 200 isoaltes of Escherichia coli,the positive rate of ESBL was 59.00%（118/200）. ESBL genotype were amplified in 105 isolates of ESBL,accounting for 88.98%. Two or more ESBL genes were determined in 11 isolates,accounting for 9.30%. CTX-M type had the highest determination rate（52.54%）. Among 118 ESBLs-producing strains,the positive detection rate of CTX-M type was the highest 52.54%. Only I class integron was determined from different positive genotype amplification bacteria. ⅠCTX-M-1 type isolate carried I class integron accounting for 72.41%,while SHV type isolate carried I class integron accounting for 50.00%. Conclusions The positive rate of ESBL of Escherichia coli in Anning is relatively high. CXT-M gene is the dominant genotype. The isolates carrying multiple ESBL genes show a gradual increase trend,and the main type is I integron.

Key words: Escherichia coli, Extended-spectrum lactamase, Genotyping, Integrase

中图分类号:

R378.2

刘存芬, 王燕, 杨溶, 胡丽香. 200株血流感染大肠埃希菌耐药性分析[J]. 检验医学, 2020, 35(12): 1263-1266.

LIU Cunfen, WANG Yan, YANG Rong, HU Lixiang. Relationship between ESBL genotyping of Escherichia coli and integron in Anning[J]. Laboratory Medicine, 2020, 35(12): 1263-1266.

图/表 4

参考文献 10

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ESBLs基因分型	序列（5'→3'）	产物长度/bp
TEM	F：TCGGGGAAAGTGCG	1 042
TEM	R：TGCTTAAATCAGTGACACC	1 042
CTX-M-1	F：TCGGTCCACTGGACAGTATG	945
CTX-M-1	R：TCCCCCGATCGTGAGTAA	945
CTX-M-8	F：CGCGAGCTAAATCAGTCCACGTTG	1 157
CTX-M-8	R：GGTTTTCCCGSGGTTACACGTGC	1 157
Ⅰ类整合子	F：CTCAAAGAGTCGTAGCCA	501
Ⅰ类整合子	R：ACTTTCAGCAGGAATTCCC	501
Ⅰ类2	F：AAGGTGTACACCGGATTCA	210
Ⅰ类2	R：GAGATAAGGAGAAGC	210
SHV	F：GAAGATCGATTGACGAATGT	869
SHV	R：TGCGGATTGGTTTTACTGAC	869
CTX-M-2	F：CAGAT（C/T）TGCTAGCTGAC（G/T）AT	856
CTX-M-2	R：CA（A/G）CATAGGTATT（A/G）AT	856
CTX-M-9	F：GCACATAGGTAGAT	862
CTX-M-9	R：GGATGAGGTAGGTG	862
Ⅰ类1	F：ATTGTAAGGTAACCT	456
Ⅰ类1	R：GCCGAGGTACTAGGGA	456
Ⅰ类3	F：GCCTTACGATGACAAAC	257
Ⅰ类3	R：GCCAGTTACCGGCATCGCT	257

ESBLs基因分型	序列（5'→3'）	产物长度/bp
TEM	F：TCGGGGAAAGTGCG	1 042
TEM	R：TGCTTAAATCAGTGACACC	1 042
CTX-M-1	F：TCGGTCCACTGGACAGTATG	945
CTX-M-1	R：TCCCCCGATCGTGAGTAA	945
CTX-M-8	F：CGCGAGCTAAATCAGTCCACGTTG	1 157
CTX-M-8	R：GGTTTTCCCGSGGTTACACGTGC	1 157
Ⅰ类整合子	F：CTCAAAGAGTCGTAGCCA	501
Ⅰ类整合子	R：ACTTTCAGCAGGAATTCCC	501
Ⅰ类2	F：AAGGTGTACACCGGATTCA	210
Ⅰ类2	R：GAGATAAGGAGAAGC	210
SHV	F：GAAGATCGATTGACGAATGT	869
SHV	R：TGCGGATTGGTTTTACTGAC	869
CTX-M-2	F：CAGAT（C/T）TGCTAGCTGAC（G/T）AT	856
CTX-M-2	R：CA（A/G）CATAGGTATT（A/G）AT	856
CTX-M-9	F：GCACATAGGTAGAT	862
CTX-M-9	R：GGATGAGGTAGGTG	862
Ⅰ类1	F：ATTGTAAGGTAACCT	456
Ⅰ类1	R：GCCGAGGTACTAGGGA	456
Ⅰ类3	F：GCCTTACGATGACAAAC	257
Ⅰ类3	R：GCCAGTTACCGGCATCGCT	257

抗菌药物	耐药		中介		敏感
抗菌药物	菌株数	耐药率/%	菌株数	中介率/%	菌株数	敏感率/%
氨曲南	125	62.40	12	8.33	60	30.00
阿米卡星	34	17.65	0	0.00	165	82.42
头孢曲松	121	60.33	32	15.63	46	23.91
头孢呋辛	142	71.13	24	12.50	65	32.31
头孢他啶	116	58.62	26	11.54	53	26.42
亚胺培南	3	33.33	0	0.00	194	97.42
庆大霉素	146	73.29	16	6.25	39	17.95
左氧氟沙星	118	59.32	35	17.14	47	23.40
复方磺胺甲噁唑	91	45.05	31	16.13	42	21.43
氨苄西林-舒巴坦	167	83.23	19	10.53	14	7.14
阿莫西林-克拉维酸	131	65.65	17	5.88	52	26.92
哌拉西林-他唑巴坦	23	13.04	0	0.00	176	88.64

抗菌药物	耐药		中介		敏感
抗菌药物	菌株数	耐药率/%	菌株数	中介率/%	菌株数	敏感率/%
氨曲南	125	62.40	12	8.33	60	30.00
阿米卡星	34	17.65	0	0.00	165	82.42
头孢曲松	121	60.33	32	15.63	46	23.91
头孢呋辛	142	71.13	24	12.50	65	32.31
头孢他啶	116	58.62	26	11.54	53	26.42
亚胺培南	3	33.33	0	0.00	194	97.42
庆大霉素	146	73.29	16	6.25	39	17.95
左氧氟沙星	118	59.32	35	17.14	47	23.40
复方磺胺甲噁唑	91	45.05	31	16.13	42	21.43
氨苄西林-舒巴坦	167	83.23	19	10.53	14	7.14
阿莫西林-克拉维酸	131	65.65	17	5.88	52	26.92
哌拉西林-他唑巴坦	23	13.04	0	0.00	176	88.64

ESBL基因分型	菌株数/株	百分比/%
TEM	27	25.71
SHV-11	11	10.48
SHV-12	7	6.67
CTX-M-1	21	20.00
CTX-M-8	18	17.14
CTX-M-9	9	8.57
CTX-M-1+TEM	4	3.81
CTX-M-1+CTX-M-9	3	2.86
CTX-M-1+CTX-M-9+TEM	3	2.86
CTX-M-9+SHV-11	1	0.95
合计	105	100.00