Carbapenemase detection and multilocus sequence typing analysis of carbapenem-resistant Enterobacteriaceae

doi:10.3969/j.issn.1673-8640.2019.07.001

Abstract

Abstract:

Objective To study the carbapenem resistance mechanism and molecular epidemiology of carbapenem-resistant Enterobacteriaceae（CRE）. Methods The identification of bacteria was performed by automatic microbial identification and analysis system. The drug susceptibility test was performed. The modified carbapenem inactivation method（mCIM） was used to screen carbapenemase,and the drug resistance genes of carbapenemase were detected by polymerase chain reaction（PCR）and gene sequencing. The molecular typing was carried out by multilocus sequence typing（MLST） analysis. Results A total of 13 carbapenem-resistant Klebsiella pneumoniae and 11 carbapenem-resistant Enterobacter cloacae were collected. The positive rate of mCIM was 100%. In the 13 carbapenem-resistant Klebsiella pneumoniae,10 isolates carried bla_KPC-2,1 isolate carried bla_NDM-1,and 1 isolate carried bla_NDM-4,1 isolate was not detected bla_KPC,bla_IMP,bla_VIM,bla_NDM and bla_OXA. The 11 carbapenem-resistant Enterobacter cloacae carried bla_NDM-1. MLST results showed that in the 13 carbapenem-resistant Klebsiella pneumoniae,12 isolates were ST11,and 1 isolate was ST571,while the 11 carbapenem-resistant Enterobacter cloacae were ST93. Conclusions The main mechanism of carbapenem-resistant Klebsiella pneumoniae is carrying bla_KPC-2,and the dominant sequence type（ST） is ST11. The main mechanism of carbapenem-resistant Enterobacter cloacae is carrying bla_NDM-1,and the dominant is ST93.

Key words: Carbapenemase, Multilocus sequence typing, Drug resistance, Klebsiella pneumoniae, Enterobacter cloacae

CLC Number:

R446.5

WEN Weihong, LIANG Yingjian, MA Furong, LI Yuzhen, XU Lingqing. Carbapenemase detection and multilocus sequence typing analysis of carbapenem-resistant Enterobacteriaceae[J]. Laboratory Medicine, 2019, 34(7): 577-582.

Figures/Tables 5

References 14

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基因名称	引物序列（5'~3'）	产物长度（bp）
KPC	F：CGGGATCCATGTCACTGTATCGCCGTC	899
	R：CGGAATTCTTACTGCCCGTTAACGCC
IMP	F：GGAATAGAGTGGCTTAAYTCTC	587
	R：TACAGCAAGCGCTGAGTAAC
NDM	F：GAATGTCTGGCAGCACACTT	480
	R：TTGGCCTTGCTGTCCTTGAT
NDM-1	F：GCTCTAGAATGGAATTGCCCAATATTAT	829
	R：CGGAATTCTCAGCGCAGCTTGTCGGCCA
VIM	F：TCTACATGACCGCGTCTGTC	748
	R：TGTGCTTGACAACGTTCGC
OXA-23-like	F：GATCGGATTGGAGAACCAGA-3′	501
	R：ATTTCTGACCGCATTTCCAT-3′
OXA-24-like	F：GGTTAGTTGGCCCCCTTAAA-3′	246
	R：AGTTGAGCGAAAAGGGGATT-3′
OXA-51-like	F：TAATGCTTTGATCGGCCTTG	353
	R：TGGATTGCACTTCATCTTGG
OXA-58-like	F：AAGTATTGGGGCTTGTGCTG	599
	R：CCCCTCTGCGCTCTACATAC

基因名称	引物序列（5'~3'）	产物长度（bp）
KPC	F：CGGGATCCATGTCACTGTATCGCCGTC	899
	R：CGGAATTCTTACTGCCCGTTAACGCC
IMP	F：GGAATAGAGTGGCTTAAYTCTC	587
	R：TACAGCAAGCGCTGAGTAAC
NDM	F：GAATGTCTGGCAGCACACTT	480
	R：TTGGCCTTGCTGTCCTTGAT
NDM-1	F：GCTCTAGAATGGAATTGCCCAATATTAT	829
	R：CGGAATTCTCAGCGCAGCTTGTCGGCCA
VIM	F：TCTACATGACCGCGTCTGTC	748
	R：TGTGCTTGACAACGTTCGC
OXA-23-like	F：GATCGGATTGGAGAACCAGA-3′	501
	R：ATTTCTGACCGCATTTCCAT-3′
OXA-24-like	F：GGTTAGTTGGCCCCCTTAAA-3′	246
	R：AGTTGAGCGAAAAGGGGATT-3′
OXA-51-like	F：TAATGCTTTGATCGGCCTTG	353
	R：TGGATTGCACTTCATCTTGG
OXA-58-like	F：AAGTATTGGGGCTTGTGCTG	599
	R：CCCCTCTGCGCTCTACATAC

菌株编号	厄他培南	亚胺培南	美罗培南	头孢噻肟	头孢他啶	头孢吡肟	哌拉西林-他唑巴坦	阿莫西林-克拉维酸	氨曲南	阿米卡星	环丙沙星	替加环素	多黏菌素
1	>64	64	>64	256	>256	128	>256	>256	>256	>256	128	2	≤0.25
2	64	8	8	>256	>256	128	>256	128	>256	8	1	2	0.5
3	>64	64	>64	>256	>256	>256	>256	>256	>256	>256	128	2	≤0.25
4	>64	32	>64	>256	>256	>256	>256	>256	>256	>256	64	2	≤0.25
5	>64	64	>64	>256	>256	>256	>256	>256	>256	>256	128	2	0.5
6	32	8	8	256	>256	64	>256	128	16	>256	128	2	0.5
7	>64	64	>64	>256	>256	>256	>256	>256	>256	>256	128	2	≤0.25
8	8	16	8	>256	>256	64	>256	256	>256	2	128	2	0.5
9	>64	64	>64	>256	>256	>256	>256	>256	>256	>256	128	2	≤0.25
10	>64	64	>64	>256	>256	>256	>256	>256	>256	>256	128	2	0.5
11	>64	64	>64	>256	>256	>256	>256	>256	>256	>256	128	2	0.5
12	32	16	4	256	>256	128	>256	>256	>256	>256	128	2	≤0.25
13	2	0.25	0.25	256	>256	64	16	64	256	>256	64	1	0.5
菌株编号	厄他培南	亚胺培南	美罗培南	头孢噻肟	头孢他啶	头孢吡肟	哌拉西林-他唑巴坦	阿莫西林-克拉维酸	氨曲南	阿米卡星	环丙沙星	替加环素	多黏菌素
14	8	16	16	256	>256	64	16	64	256	>256	64	1	0.5
15	>64	16	16	>256	>256	>256	>256	256	>256	>256	128	2	0.5
16	32	16	8	>256	>256	256	>256	256	>256	>256	128	2	0.5
17	8	8	8	>256	>256	128	>256	256	>256	2	64	2	≤0.25
18	32	4	8	>256	>256	256	>256	256	>256	>256	128	2	0.5
19	32	16	8	256	>256	64	64	256	128	>256	64	2	0.5
20	32	8	8	>256	>256	64	>256	256	>256	>256	64	2	≤0.25
21	8	16	16	>256	>256	64	>256	256	>256	>256	128	2	1
22	8	16	8	>256	>256	64	>256	256	>256	2	64	2	≤0.25
23	16	16	8	>256	>256	64	>256	256	>256	2	64	1	≤0.25
24	8	32	16	>256	>256	128	>256	256	>256	2	64	2	≤0.25

菌株编号	厄他培南	亚胺培南	美罗培南	头孢噻肟	头孢他啶	头孢吡肟	哌拉西林-他唑巴坦	阿莫西林-克拉维酸	氨曲南	阿米卡星	环丙沙星	替加环素	多黏菌素
1	>64	64	>64	256	>256	128	>256	>256	>256	>256	128	2	≤0.25
2	64	8	8	>256	>256	128	>256	128	>256	8	1	2	0.5
3	>64	64	>64	>256	>256	>256	>256	>256	>256	>256	128	2	≤0.25
4	>64	32	>64	>256	>256	>256	>256	>256	>256	>256	64	2	≤0.25
5	>64	64	>64	>256	>256	>256	>256	>256	>256	>256	128	2	0.5
6	32	8	8	256	>256	64	>256	128	16	>256	128	2	0.5
7	>64	64	>64	>256	>256	>256	>256	>256	>256	>256	128	2	≤0.25
8	8	16	8	>256	>256	64	>256	256	>256	2	128	2	0.5
9	>64	64	>64	>256	>256	>256	>256	>256	>256	>256	128	2	≤0.25
10	>64	64	>64	>256	>256	>256	>256	>256	>256	>256	128	2	0.5
11	>64	64	>64	>256	>256	>256	>256	>256	>256	>256	128	2	0.5
12	32	16	4	256	>256	128	>256	>256	>256	>256	128	2	≤0.25
13	2	0.25	0.25	256	>256	64	16	64	256	>256	64	1	0.5
菌株编号	厄他培南	亚胺培南	美罗培南	头孢噻肟	头孢他啶	头孢吡肟	哌拉西林-他唑巴坦	阿莫西林-克拉维酸	氨曲南	阿米卡星	环丙沙星	替加环素	多黏菌素
14	8	16	16	256	>256	64	16	64	256	>256	64	1	0.5
15	>64	16	16	>256	>256	>256	>256	256	>256	>256	128	2	0.5
16	32	16	8	>256	>256	256	>256	256	>256	>256	128	2	0.5
17	8	8	8	>256	>256	128	>256	256	>256	2	64	2	≤0.25
18	32	4	8	>256	>256	256	>256	256	>256	>256	128	2	0.5
19	32	16	8	256	>256	64	64	256	128	>256	64	2	0.5
20	32	8	8	>256	>256	64	>256	256	>256	>256	64	2	≤0.25
21	8	16	16	>256	>256	64	>256	256	>256	>256	128	2	1
22	8	16	8	>256	>256	64	>256	256	>256	2	64	2	≤0.25
23	16	16	8	>256	>256	64	>256	256	>256	2	64	1	≤0.25
24	8	32	16	>256	>256	128	>256	256	>256	2	64	2	≤0.25

菌株编号	科室来源	样本类型	分离日期	疾病	耐药基因	ST
1	重症监护病房3区	痰液	2016-01-22	呼吸衰竭	bla_KPC-2	11
2	脑科1区	中段尿	2016-04-01	腹痛查因	bla_NDM-1	571
3	脑科1区	中段尿	2016-04-24	颅内血肿	bla_KPC-2	11
4	重症监护病房2区	痰液	2016-06-23	脑梗塞	bla_KPC-2	11
5	呼吸内科2区	痰液	2016-06-23	肺结核	bla_KPC-2	11
6	泌尿外科	中段尿	2016-07-06	左肾结石	bla_NDM-4	11
7	脑科5区	中段尿	2016-08-05	脑出血	bla_KPC-2	11
8	肾内科	中段尿	2016-08-27	梗阻性肾病	bla_KPC-2	11
9	脑科5区	中段尿	2016-11-24	脑出血	bla_KPC-2	11
10	呼吸内科2区	痰液	2016-12-07	脑梗塞	bla_KPC-2	11
11	脑科5区	中段尿	2016-12-15	脑出血	bla_KPC-2	11
12	脑科5区	中段尿	2017-02-10	脑出血	bla_KPC-2	11
13	脑科1区	中段尿	2017-02-22	脑外伤		11
14	脑科5区	中段尿	2015-08-21	脑血管意外	bla_NDM-1	93
15	脑科1区	中段尿	2015-10-14	脑挫裂伤	bla_NDM-1	93
16	脑科1区	中段尿	2015-11-08	脑挫裂伤	bla_NDM-1	93
17	脑科5区	中段尿	2015-11-09	脑血管意外	bla_NDM-1	93
18	脑科5区	中段尿	2015-12-06	脑出血	bla_NDM-1	93
19	脑科1区	中段尿	2015-12-19	脑挫裂伤	bla_NDM-1	93
20	脑科5区	中段尿	2016-01-22	脑血管意外	bla_NDM-1	93
21	脑科1区	中段尿	2016-01-23	脑出血	bla_NDM-1	93
22	脑科5区	中段尿	2016-04-22	脑出血	bla_NDM-1	93
23	脑科5区	中段尿	2016-09-24	脑挫裂伤	bla_NDM-1	93
24	脑科5区	中段尿	2016-11-21	脑出血	bla_NDM-1	93