北京市某院难辨梭菌临床分离株耐药性及毒素基因携带情况分析

doi:10.3969/j.issn.1673-8640.2020.09.002

摘要/Abstract

摘要：

目的了解航天中心医院难辨梭菌临床分离株的耐药性及毒素基因携带情况。方法收集不明原因腹泻患者的粪便样本335例,采用TaqMan-MGB探针实时荧光定量聚合酶链反应（PCR）进行菌属鉴定及毒素基因检测。对tpi基因阳性样本进行厌氧培养、分离及药物敏感性试验。回顾性分析分离出A⁺B⁺菌株和A^-B⁺菌株患者的临床资料。结果 335例粪便样本中,56例（16.7%）检出难辨梭菌tpi基因,其中10株（17.9%）A^-B⁺菌株、46株（82.1%）A⁺B⁺菌株。检出菌株对阿莫西林、甲硝唑、克林霉素、万古霉素、莫西沙星、氨苄西林-舒巴坦及美罗培南的耐药率分别为7.1%、10.7%、75.0%、0、57.1%、5.4%、8.9%。有29株（51.8%）菌对克林霉素、莫西沙星双重耐药。分离出2种菌的患者病史、临床用药史、常规实验室检测指标差异均无统计学意义（P>0.05）。结论航天中心医院难辨梭菌临床分离株以多重耐药模式多见,A⁺B⁺菌株和A^-B⁺菌株感染患者临床特征无差异。

关键词: 难辨梭菌, 药物敏感性, 毒素基因

Abstract:

Objective To understand the drug resistance and toxin gene carrying status of Clostridium difficile clinical isolates in Aerospace Center Hospital. Methods The stool specimens of clinical unexplained diarrhea patients were detected by TaqMan-MGB probe real-time fluorescence quantitative polymerase chain reaction（PCR） to identify genus gene and toxin gene. Anaerobic culture,isolation and drug susceptibility test were conducted for the positive specimens of tpi gene. The clinical records were analyzed retrospectively,in order to compare the difference between A⁺B⁺ isolates and A^-B⁺ isolates. Results The tpi gene of 56 specimens was positive in 335 stool specimens,the positive rate was 16.7%,in which 10（17.9%） isolates were A^-B⁺ isolates,and 46（82.1%） isolates were A⁺B⁺ isolates. The drug resistance rates to amoxicillin,metronidazole,clindamycin,vancomycin,moxifloxacin,ampicillin-sulbactam and meropenem were 7.1%,10.7%,75.0%,0,57.1%,5.4% and 8.9%,respectively. There were 29（51.8%） isolates with drug resistance to both clindamycin and moxifloxacin. There was no statistical significance in patient history,clinical drug history and laboratory data（P>0.05）. Conclusions The multiple drug resistance of Clostridium difficile in Aerospace Center Hospital is common. The clinical characteristics are not different between A⁺B⁺ isolates and A^-B⁺ isolates.

Key words: Clostridium difficile, Drug susceptibility, Toxin gene

中图分类号:

R378.8

邵冬华, 宋燕, 梁国威. 北京市某院难辨梭菌临床分离株耐药性及毒素基因携带情况分析[J]. 检验医学, 2020, 35(9): 864-867.

SHAO Donghua, SONG Yan, LIANG Guowei. Analysis on drug resistance and toxin gene carrying status of Clostridium difficile clinical isolates in a hospital of Beijing[J]. Laboratory Medicine, 2020, 35(9): 864-867.

图/表 7

参考文献 13

[1]	MILLER B A,CHEN L F,SEXTON D J,et al.Comparison of the burdens of hospital-onset,healthcare facility-associated Clostridium difficile infection and of healthcare-associated infection due to methicillin-resistant Staphylococcus aureus in community hospitals[J]. Infect Control Hosp Epidemiol,2011,32(4):387-390.
[2]	SHIN B M,KUAK E Y,YOO S J,et al.Emerging toxin A-B+ variant strain of Clostridium difficile responsible for pseudomembranous colitis at a tertiary care hospital in Korea[J]. Diagn Microbiol Infect Dis,2008,60(4):333-337.
[3]	邵冬华,季娜,梁国威,等. TaqMan-MGB探针实时荧光定量PCR联合检测难辨梭菌菌属基因及毒素基因方法学的建立[J]. 中华流行病学杂志,2014,35(5):576-580.
[4]	MCDONALD L C,GERDING D N,JOHNSON S,et al.Clinical practice guidelines for Clostridium difficile infection in adults and children:2017 update by the Infectious Diseases Society of America(IDSA) and Society for Healthcare Epidemiology of America(SHEA)[J]. Clin Infect Dis,2018,66(7):e1-e48.
[5]	DHALLUIN A,LEMÉE L,PESTEL-CARON M,et al. Genotypic differentiation of twelve Clostridium species by polymorphism analysis of the triosephosphate isomerase(tpi) gene[J]. Syst Appl Microbiol,2003,26(1):90-96.
[6]	BAUER M P,NOTERMANS D W,VAN BENTHEM B H,et al. Clostridium difficile infection in Europe:a hospital-based survey[J]. Lancet,2011,377(9759):63-73.
[7]	NGAMSKULRUNGROJ P,SANMEE S,PUTSATHIT P,et al.Molecular epidemiology of Clostridium difficile infection in a large teaching hospital in Thailand[J]. PLoS One,2015,10(7):e0134771.
[8]	KUWATA Y,TANIMOTO S,SAWABE E,et al.Molecular epidemiology and antimicrobial susceptibility of Clostridium difficile isolated from a university teaching hospital in Japan[J]. Eur J Clin Microbiol Infect Dis,2015,34(4):763-772.
[9]	SHIN B M,KUAK E Y,YOO S J,et al.Emerging toxin A-B+ variant strain of Clostridium difficile responsible for pseudomembranous colitis at a tertiary care hospital in Korea[J]. Diagn Microbiol Infect Dis,2008,60(4):333-337.
[10]	KIM J,PAI H,SEO M R,et al.Clinical and microbiologic characteristics of tcdA-negative variant Clostridium difficile infections[J]. BMC Infect Dis,2012,12:109.
[11]	TICKLER I A,GOERING R V,WHITMORE J D,et al.Strain types and antimicrobial resistance patterns of Clostridium difficile isolates from the United States,2011 to 2013[J]. Antimicrol Agents Chemother,2014,58(7):4214-4218.
[12]	TKHAWKHO L,NITZAN O,PASTUKH N,et al.Antimicrobial susceptibility of Clostridium difficile isolates in Israel[J]. J Glob Antimicrob Resist,2017,10:161-164.
[13]	FRAGA E G,NICODEMO A C,SAMPAIO J L.Antimicrobial susceptibility of Brazilian Clostridium difficile strains determined by agar dilution and disk diffusion[J]. Braz J Infect Dis,2016,20(5):476-481.

引物名称	序列（5'~3'）	序列位置（基因Bank号：NC_013315）
tpi	TTCAAATGTAGCCGAAATAATGG	3531170-3531192
	GAAATTAACAAGGTCTAGGTAATCATTAC	3531241-3531269
tpi-probe	FAM-ATATAGATGGAGCTTTAGTA-MGB	3531202-3531221
tcdA	ACACCTTAACCCAGCCATAGAGT	720522-720544
	TTTCTGCGGTAGCTGAATTAAATA	720590-720613
tcdA-probe	FAM-TAACTTCACAGATACTACTA-MGB	720552-720571
tcdB	TAAATCAATGGAAAGATGTAAATAGTG	711080-711106
	TTTTTTCAATGTGTTTATCAA AAATGC	711139-711165
tcdB-probe	FAM-CATAAAAAACATTAACATTATA-MGB	711109-711130
tcdAT	GATATTACTTCGAGCCTAATACAGC	727034-727058
	CTATCTGAGGTAAACCATTTCTAAAGT	727103-727129
tcdAT-probe	FAM-TGCGAATGGTTATAAAA-MGB	727065-727081

引物名称	序列（5'~3'）	序列位置（基因Bank号：NC_013315）
tpi	TTCAAATGTAGCCGAAATAATGG	3531170-3531192
	GAAATTAACAAGGTCTAGGTAATCATTAC	3531241-3531269
tpi-probe	FAM-ATATAGATGGAGCTTTAGTA-MGB	3531202-3531221
tcdA	ACACCTTAACCCAGCCATAGAGT	720522-720544
	TTTCTGCGGTAGCTGAATTAAATA	720590-720613
tcdA-probe	FAM-TAACTTCACAGATACTACTA-MGB	720552-720571
tcdB	TAAATCAATGGAAAGATGTAAATAGTG	711080-711106
	TTTTTTCAATGTGTTTATCAA AAATGC	711139-711165
tcdB-probe	FAM-CATAAAAAACATTAACATTATA-MGB	711109-711130
tcdAT	GATATTACTTCGAGCCTAATACAGC	727034-727058
	CTATCTGAGGTAAACCATTTCTAAAGT	727103-727129
tcdAT-probe	FAM-TGCGAATGGTTATAAAA-MGB	727065-727081

耐药组合	菌株数	百分比/%
克林霉素+莫西沙星	29	51.8
克林霉素+莫西沙星+甲硝唑	5	8.9
甲硝唑+克林霉素+莫西沙星+ 美罗培南	4	7.1
阿莫西林+甲硝唑+克林霉素+ 莫西沙星+氨苄西林-舒巴坦	2	3.6

耐药组合	菌株数	百分比/%
克林霉素+莫西沙星	29	51.8
克林霉素+莫西沙星+甲硝唑	5	8.9
甲硝唑+克林霉素+莫西沙星+ 美罗培南	4	7.1
阿莫西林+甲硝唑+克林霉素+ 莫西沙星+氨苄西林-舒巴坦	2	3.6

菌株	心血管疾病	恶性肿瘤	肝胆疾病	胃肠道疾病	肾脏疾病	糖尿病
A⁺B⁺菌株	32（70）	13（28）	11（24）	26（56）	7（15）	14（30）
A^-B⁺菌株	8（80）	3（30）	2（20）	3（30）	4（40）	5（50）
P值	0.707	1.000	1.000	0.171	0.093	0.281