南京某医院成人耐甲氧西林金黄色葡萄球菌检出情况及耐药基因分析

doi:10.3969/j.issn.1673-8640.2019.03.006

摘要/Abstract

摘要：

目的了解江苏省中西医结合医院2011—2017年成人耐甲氧西林金黄色葡萄球菌（MRSA）临床分离株的检出情况及其耐药基因,为临床合理用药提供参考依据。方法采用美国临床实验室标准化协会（CLSI）推荐方法鉴定MRSA。采用纸片扩散法对MRSA菌株进行药物敏感性试验。采用聚合酶链反应（PCR）检测β-内酰胺类耐药基因mecA,氨基糖苷修饰酶基因aac（6′）/aph（2″）、aph（3）-Ⅲ,大环内酯类23s rRNA甲基化酶基因ermA/C,四环素类核糖体保护蛋白基因tetM/K,喹诺酮类耐药基因qnrA/B和耐消毒剂基因qacA/B。结果 MRSA的总检出率为31.95%,主要分离自痰液和创口分泌物样本;检出率居前3位的科室依次为呼吸科、外科和重症监护病房。未检出对万古霉素、利奈唑胺和替加环素耐药的MRSA菌株;仅有0.49%的MRSA对奎奴普汀耐药,对复方磺胺甲噁唑、利福平、环丙沙星、四环素、庆大霉素、克林霉素、红霉素的耐药率分别为11.18%、11.57%、21.47%、63.63%、44.31%、59.41%、62.35%。多重耐药基因mecA阳性率为100%,aac（6′）/aph（2″）、aph（3）-Ⅲ、ermA/C、tetM/K、qnrA/B、qacA/B基因检出率分别为36.67%、27.94%、68.43%、57.35%、23.63%和52.45%。结论 2011—2017年江苏省中西医结合医院成人MRSA多重耐药基因检出率高。MRSA对多种抗菌药物都不同程度耐药,临床应以药物敏感性试验结果为合理用药的最主要依据。

关键词: 耐甲氧西林金黄色葡萄球菌, 多重耐药, 基因

Abstract:

Objective To investigate the determination and drug resistance gene of methicillin-resistant Staphylococcus aureus （MRSA） isolated from adults in Jiangsu Province Hospital on Integration of Chinese and Western Medicine from 2011 to 2017,and to provide a reference for clinical rational drug use. Methods According to the Clinical and Laboratory Standards Institute （CLSI） guidelines,MRSA were identified. The drug susceptibility was determined by disk diffusion method. The β-lactam resistance gene mecA,aminoglycoside modifying enzyme genes aac（6′）/aph（2″）and aph（3）-Ⅲ,macrolide 23s rRNA methylase gene ermA/C,tetracycline-like ribosomal protective protein gene tetM/K,quinolone resistance gene qnrA/B and disinfectant resistance gene qacA/B were determined by polymerase chain reaction （PCR）. Results The determination rate of MRSA was 31.95%,and the isolates of MRSA were mainly isolated from sputum and wound secretion samples. The top 3 departments were respiratory department,surgery department and intensive care unit. There was no MRSA isolate being resistant to vancomycin,linezolid and tigecycline. There were 0.49% MRSA isolates being resistant to quinupristin. The drug resistance rates to cotrimoxazole,rifampicin,ciprofloxacin,tetracycline,gentamicin,clindamycin and erythromycin were 11.18%,11.57%,21.47%,63.63%,44.31%,59.41% and 62.35%,respectively. The determination rate of mecA was 100%. The determination rates of aac（6′）/aph（2″）,aph（3）-Ⅲ,ermA/C,tetM/K,qnrA/B and qacA/B were 36.67 %,27.94%,68.43%,57.35%,23.63% and 52.45%,respectively.Conclusions From 2011 to 2017,the determination rate of MRAS multidrug resistance gene is high. MRSA shows various degrees of drug resistance to many antibiotics. Drug susceptibility test's results can be used reference for clinical medication.

Key words: Methicillin-resistant Staphylococcus aureus, Multidrug resistance, Gene

中图分类号:

R446.62

张利, 孙林春. 南京某医院成人耐甲氧西林金黄色葡萄球菌检出情况及耐药基因分析[J]. 检验医学, 2019, 34(3): 215-220.

ZHANG Li, SUN Linchun. Determination and drug resistance gene of methicillin-resistant Staphylococcus aureus isolated from adults in a hospital of Nanjing[J]. Laboratory Medicine, 2019, 34(3): 215-220.

图/表 6

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基因名称	引物序列（5'~3'）		长度（bp）
mecA	上游：AAAATCGATGGTAAAGGTTGGC;	下游：AGTTCTGCAGTACCGGATTTGC	533
aac（6′）/aph（2″）	上游：CCAAGAGCAATAAGGGCATA;	下游：CACTATCATAACCACTACCG	220
Aph（3）-Ⅲ	上游：GCCGATGTGGATTGCGAAAA;	下游：GCTTGATCCCCAGTAAGTCA	292
ermA/C	上游：GTTCAAGAACAATCACAGAG;	下游：GGATCAGGAAAAGGACATTTTAC	533
tetM/K	上游：GTGTGACGAACTTTACCGAA;	下游：GCTTTCTATCTCCAAGAACAC	501
qacA/B	上游：GGTTGTGGAAGAACTTTCTCCTT;	下游：CCAATTCCGGAAGGTAACAC	416
qnrA/B	上游：ATTTCTCACGCCAGGATTTG;	下游：GATCGGCAAAGGTTAGGTCA	515

基因名称	引物序列（5'~3'）		长度（bp）
mecA	上游：AAAATCGATGGTAAAGGTTGGC;	下游：AGTTCTGCAGTACCGGATTTGC	533
aac（6′）/aph（2″）	上游：CCAAGAGCAATAAGGGCATA;	下游：CACTATCATAACCACTACCG	220
Aph（3）-Ⅲ	上游：GCCGATGTGGATTGCGAAAA;	下游：GCTTGATCCCCAGTAAGTCA	292
ermA/C	上游：GTTCAAGAACAATCACAGAG;	下游：GGATCAGGAAAAGGACATTTTAC	533
tetM/K	上游：GTGTGACGAACTTTACCGAA;	下游：GCTTTCTATCTCCAAGAACAC	501
qacA/B	上游：GGTTGTGGAAGAACTTTCTCCTT;	下游：CCAATTCCGGAAGGTAACAC	416
qnrA/B	上游：ATTTCTCACGCCAGGATTTG;	下游：GATCGGCAAAGGTTAGGTCA	515

年份	SA（株）	MRSA（株）	MRSA检出率（%）
2011	451	125	27.72
2012	431	111	25.75
2013	445	141	31.69
2014	435	154	35.40^*
2015	453	158	34.88^*
2016	486	164	33.74^*
2017	491	167	33.40^*

年份	SA（株）	MRSA（株）	MRSA检出率（%）
2011	451	125	27.72
2012	431	111	25.75
2013	445	141	31.69
2014	435	154	35.40^*
2015	453	158	34.88^*
2016	486	164	33.74^*
2017	491	167	33.40^*

样本类型	SA（株）	MRSA（株）	MRSA检出率（%）
痰液	1 246	608	48.80^**
创口分泌物	496	187	37.70^*
血液	1 012	152	15.02^**
前列腺液	144	17	11.81^**
穿刺液	98	27	27.55
引流物	65	16	24.62
胸腹腔积液	131	13	9.92^**
合计	3 192	1 020	31.95