耐碳青霉烯类肠杆菌科细菌的耐药机制探讨

doi:10.3969/j.issn.1673-8640.2017.12.002

摘要/Abstract

摘要：

目的探讨耐碳青霉烯类肠杆菌科细菌的耐药机制。方法收集上海交通大学附属第六人民医院2011年1月—2013年12月临床分离到的80株碳青霉烯类抗菌药物耐药的肠杆菌科细菌,采用Vitek Compao60鉴定系统进行细菌鉴定和药物敏感性检测,用纸片扩散法检测细菌对亚胺培南和美罗培南的敏感性,用改良Hodge试验、抑制剂的双纸片增效试验检测碳青霉烯酶表型,用聚合酶链反应（PCR）检测bla_KPC、bla_IMP、bla_OXA₂₃、bla_OXA₅₁、bla_OXA₄₈、bla_VIM、bla_NMD。结果 80株菌株对常见多种抗菌药物耐药;改良Hodge试验和硼酸抑制试验与KPC酶检测的一致率分别为47.5%和97.5%;bla_KPC、bla_IMP、bla_OXA₂₃、bla_OXA₅₁、bla_OXA₄₈、bla_VIM和bla_NMD的检出率分别为88.75%、5.00%、3.75%、3.75%、0.00%、0.00%和0.00%,所有基因均阴性的检出率为5.00%。结论上海交通大学附属第六人民医院肠杆菌科细菌耐碳青霉烯类抗菌药物的主要机制是携带KPC-2型碳青霉烯酶,硼酸抑制试验能快速、准确地检测出KPC酶。

关键词: 肠杆菌科细菌, 碳青霉烯酶, 改良Hodge试验, 双纸片增效试验, KPC

Abstract:

Objective To study the mechanism of carbapenems-resistant Enterobacteriaceae. Methods A total of 80 isolates of Enterobacteriaceae being resistant to carbapenems were isolated from patients in Shanghai Sixth People's Hospital from January 2011 to December 2013. The isolates were identified by Vitek Compao60 automatic system,and antimicrobial susceptibility was determined as well. The susceptibilities to imipenem and meropenem were determined Kirby-Bauer disc diffusion method. The modified Hodge test and inhibitor-potentiated double-disc diffusion test were performed for determining carbapenemase phenotype. Polymerase chain reaction（PCR） was performed for determining bla_KPC,bla_IMP,bla_OXA₂₃,bla_OXA₅₁,bla_OXA₄₈,bla_VIM and bla_NMD. Results The isolates were resistant to many antibiotics. The consistencies of modified Hodge test and aminophenylboronic acid inhibition test to KPC were 47.5% and 97.5%. The determination rates of bla_KPC,bla_IMP,bla_OXA₂₃,bla_OXA₅₁,bla_OXA₄₈,bla_VIM and bla_NMD were 88.75%,5.00%,3.75%,3.75%,0.00%,0.00% and 0.00%. The determination rate of isolates being negative for all the tested genes was 5.00%. Conclusions KPC-2 carbapenemase is the main mechanism of carbapenems-resistant Enterobacteriaceae in Shanghai Sixth People's Hospital. The inhibitor-potentiated double-disc diffusion test can determine KPC easily and correctly.

Key words: Enterobacteriaceae, Carbapenemase, Modified Hodge test, Inhibitor-potentiated double-disc diffusion test, KPC

中图分类号:

R446.5

王坚镪, 吴琼, 庄亦晖, 陈敏, 汤瑾. 耐碳青霉烯类肠杆菌科细菌的耐药机制探讨[J]. 检验医学, 2017, 32(12): 1080-1084.

WANG Jianqiang, WU Qiong, ZHUANG Yihui, CHEN Min, TANG Jin. Mechanism of carbapenems-resistant Enterobacteriaceae[J]. Laboratory Medicine, 2017, 32(12): 1080-1084.

图/表 4

参考文献 15

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序列	底物	序列（5'~3'）	大小（bp）	退火温度（℃）
KPC	F_KPC	TCTTGCTGCCGCTGTGCTGG	654	56
	R_KPC	CCGTTGACGCCCAATCCCTC
IMP	F_IMP	TGAGCAAGTTATCTGTATTC	773	55
	R_IMP	TTAGTTGCTTGGTTTTGATG
OXA23	F_OXA23	GTGTCATAGTATTCGTCGTTAG	592	60
	R_OXA23	TATCAACCTGCTGTCCAAT
OXA48	F_OXA48	TTGGTGGCATCGATTATCGG	744	62
	R_OXA48	GAGCACTTCTTTTGTGATGGC
OXA51	F_OXA51	GTCATAGTATTCGTCGTTAGA	301	60
	R_OXA51	GCACGAGCAAGATCATTACCATAGC
VIM	F_VIM	TTATGGAGCAGCAACCGATGT	920	60
	R_VIM	CAAAAGTCCCGCTCCAACGA
NMD	F_ NMD	GCCATGTCACTGAATACTCGT	350	60
	R_ NMD	GCGATCCTTCCAACTCGT

序列	底物	序列（5'~3'）	大小（bp）	退火温度（℃）
KPC	F_KPC	TCTTGCTGCCGCTGTGCTGG	654	56
	R_KPC	CCGTTGACGCCCAATCCCTC
IMP	F_IMP	TGAGCAAGTTATCTGTATTC	773	55
	R_IMP	TTAGTTGCTTGGTTTTGATG
OXA23	F_OXA23	GTGTCATAGTATTCGTCGTTAG	592	60
	R_OXA23	TATCAACCTGCTGTCCAAT
OXA48	F_OXA48	TTGGTGGCATCGATTATCGG	744	62
	R_OXA48	GAGCACTTCTTTTGTGATGGC
OXA51	F_OXA51	GTCATAGTATTCGTCGTTAGA	301	60
	R_OXA51	GCACGAGCAAGATCATTACCATAGC
VIM	F_VIM	TTATGGAGCAGCAACCGATGT	920	60
	R_VIM	CAAAAGTCCCGCTCCAACGA
NMD	F_ NMD	GCCATGTCACTGAATACTCGT	350	60
	R_ NMD	GCGATCCTTCCAACTCGT

抗菌药物	耐药	中介	敏感
亚胺培南	100.00	0.00	0.00
美罗培南	100.00	0.00	0.00
阿米卡星	76.25	0.00	23.75
庆大霉素	80.00	0.00	20.00
妥布霉素	83.75	1.25	15.00
头孢噻肟	100.00	0.00	0.00
头孢他啶	100.00	0.00	0.00
头孢吡肟	100.00	0.00	0.00
头孢哌酮-舒巴坦	100.00	0.00	0.00
左氧氟沙星	98.75	0.00	1.25
替加环素	0.00	57.50	42.50

抗菌药物	耐药	中介	敏感
亚胺培南	100.00	0.00	0.00
美罗培南	100.00	0.00	0.00
阿米卡星	76.25	0.00	23.75
庆大霉素	80.00	0.00	20.00
妥布霉素	83.75	1.25	15.00
头孢噻肟	100.00	0.00	0.00
头孢他啶	100.00	0.00	0.00
头孢吡肟	100.00	0.00	0.00
头孢哌酮-舒巴坦	100.00	0.00	0.00
左氧氟沙星	98.75	0.00	1.25
替加环素	0.00	57.50	42.50