质粒介导的革兰阴性杆菌磷霉素耐药基因的研究

doi:10.3969/j.issn.1673-8640.2015.06.019

摘要/Abstract

摘要： 目的

了解质粒介导的磷霉素耐药基因在革兰阴性杆菌中的流行情况,并分析其与耐药性的关系。

方法

收集临床分离的133株革兰阴性杆菌,其中大肠埃希菌52株(39.1%)、铜绿假单胞菌36株(27.1%)、肺炎克雷伯菌31株(23.3%)、鲍曼不动杆菌14株(10.5%)。采用K-B纸片法进行药敏试验,通过聚合酶链反应(PCR)检测质粒介导的磷霉素耐药基因fosA、fosA3、fosC2,分析其与耐药性的关系。

结果

大肠埃希菌对磷霉素的耐药率为9.6%(5/52),铜绿假单胞菌、肺炎克雷伯菌和鲍曼不动杆菌的耐药率分别为36.1%(13/36)、74.2%(23/31)和64.3%(9/14)。磷霉素耐药基因fosA、fosA3阳性各5株,均为大肠埃希菌或肺炎克雷伯菌,未检出fosC2基因。10株质粒介导磷霉素耐药基因阳性菌株中,7株对磷霉素表现出耐药,其余3株表现为敏感。

结论

铜绿假单胞菌、肺炎克雷伯菌、鲍曼不动杆菌对磷霉素的耐药率均远高于大肠埃希菌。大肠埃希菌和肺炎克雷伯菌发现有fosA和fosA3基因,铜绿假单胞菌和鲍曼不动杆菌未见,尚不能解释表型试验结果,因此有必要进行更全面的耐药基因研究。

关键词: 革兰阴性杆菌, 质粒, 磷霉素, 耐药基因

Abstract: Objective

To investigate the prevalence of plasmid-mediated fosfomycin resistance genes in Gram-negative bacilli and analyze the correlation with drug resistance.

Methods

A total of 133 Gram-negative bacilli were collected, including Escherichia coli (52 isolates, 39.1%), Pseudomonas aeruginosa (36 isolates, 27.1%), Klebsiella pneumoniae (31 isolates, 23.3%) and Acinetobacter baumannii (14 isolates, 10.5%). The antimicrobial susceptibility test was performed by Kirby-Bauer (K-B) method, and the plasmid-mediated fosfomycin resistance genes(fosA, fosA3 and fosC2) were detected by polymerase chain reaction (PCR).

Results

The fosfomycin-resistant rate of Escherichia coli was 9.6%(5/52), and the ones of Pseudomonas aeruginosa, Klebsiella pneumoniae and Acinetobacter baumannii were 36.1%(13/36), 74.2%(23/31) and 64.3%(9/14), respectively. Five of 133 isolates were positive for fosA, and 5 isolates were positive for fosA3, all belonging to Escherichia coli or Klebsiella pneumoniae, while there was no fosC2 gene. Seven of 10 isolates with plasmid-mediated fosfomycin resistance genes were resistant to fosfomycin, and the other 3 isolates were sensitive.

Conclusions

The fosfomycin-resistant rates of Pseudomonas aeruginosa, Klebsiella pneumoniae and Acinetobacter baumannii are higher than that of Escherichia coli. The fosA and fosA3 genes are detected in Escherichia coli and Klebsiella pneumoniae, whereas no fos gene is detected in Pseudomonas aeruginosa and Acinetobacter baumannii, highlighting the necessity of a comprehensive research in regard to fosfomycin resistance genes.

Key words: Gram-negative bacillus, Plasmid, Fosfomycin, Resistance gene

中图分类号:

R446.5

蔡红妹, 唐瑜, 蒋晓飞. 质粒介导的革兰阴性杆菌磷霉素耐药基因的研究[J]. 检验医学, 2015, 30(6): 622-625.

CAI Hongmei, TANG Yu, JIANG Xiaofei. Plasmid-mediated fosfomycin resistance genes in Gram-negative bacilli[J]. Laboratory Medicine, 2015, 30(6): 622-625.

图/表 3

参考文献 14

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引物名称	引物序列(5'-3')	产物大小(bp)
fosC2-F fosC2-R	TGGAGGCTACTTGGATTTG AGGCTACCGCTATGGATTT	217
fosA-F fosA-R	ATCTGTGGGTCTGCCTGTCGT ATGCCCGCATAGGGCTTCT	271
fosA3-F fosA3-R	GCGTCAAGCCTGGCATTT GCCGTCAGGGTCGAGAAA	282

引物名称	引物序列(5'-3')	产物大小(bp)
fosC2-F fosC2-R	TGGAGGCTACTTGGATTTG AGGCTACCGCTATGGATTT	217
fosA-F fosA-R	ATCTGTGGGTCTGCCTGTCGT ATGCCCGCATAGGGCTTCT	271
fosA3-F fosA3-R	GCGTCAAGCCTGGCATTT GCCGTCAGGGTCGAGAAA	282

抗菌药物	大肠埃希菌 (52株)		铜绿假单胞菌 (36株)		肺炎克雷伯菌 (31株)		鲍曼不动杆菌 (14株)		合计 (133株)
抗菌药物	耐药菌株	耐药率 (%)	耐药菌株	耐药率 (%)	耐药菌株	耐药率 (%)	耐药菌株	耐药率 (%)	耐药菌株	耐药率 (%)
氨苄西林	38	73.1	26	72.2	31	100.0	8	57.1	103	77.4
亚胺培南	1	1.9	13	36.1	1	3.2	5	35.7	20	15.0
美罗培南	2	3.8	12	33.3	1	3.2	4	28.6	19	14.3
阿米卡星	5	9.6	8	22.2	4	12.9	2	14.3	19	14.3
庆大霉素	22	42.3	14	38.9	5	16.1	4	28.6	45	33.8
妥布霉素	20	38.5	9	25.0	6	19.4	4	28.6	39	29.3
环丙沙星	23	44.2	16	44.4	9	29.0	5	35.7	53	39.8
磷霉素^*	5	9.6	13	36.1	23	74.2	9	64.3	50	37.6
头孢他啶	24	46.2	7	19.4	10	32.3	4	28.6	45	33.8
头孢吡肟	21	40.4	5	13.9	9	29.0	4	28.6	39	29.3
哌拉西林	36	69.2	7	19.4	14	45.2	4	28.6	61	45.9
哌拉西林/他唑巴坦	3	5.8	4	11.1	4	12.9	4	28.6	15	11.3

抗菌药物	大肠埃希菌 (52株)		铜绿假单胞菌 (36株)		肺炎克雷伯菌 (31株)		鲍曼不动杆菌 (14株)		合计 (133株)
抗菌药物	耐药菌株	耐药率 (%)	耐药菌株	耐药率 (%)	耐药菌株	耐药率 (%)	耐药菌株	耐药率 (%)	耐药菌株	耐药率 (%)
氨苄西林	38	73.1	26	72.2	31	100.0	8	57.1	103	77.4
亚胺培南	1	1.9	13	36.1	1	3.2	5	35.7	20	15.0
美罗培南	2	3.8	12	33.3	1	3.2	4	28.6	19	14.3
阿米卡星	5	9.6	8	22.2	4	12.9	2	14.3	19	14.3
庆大霉素	22	42.3	14	38.9	5	16.1	4	28.6	45	33.8
妥布霉素	20	38.5	9	25.0	6	19.4	4	28.6	39	29.3
环丙沙星	23	44.2	16	44.4	9	29.0	5	35.7	53	39.8
磷霉素^*	5	9.6	13	36.1	23	74.2	9	64.3	50	37.6
头孢他啶	24	46.2	7	19.4	10	32.3	4	28.6	45	33.8
头孢吡肟	21	40.4	5	13.9	9	29.0	4	28.6	39	29.3
哌拉西林	36	69.2	7	19.4	14	45.2	4	28.6	61	45.9
哌拉西林/他唑巴坦	3	5.8	4	11.1	4	12.9	4	28.6	15	11.3

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