MALDI-TOF MS技术对VIM型和SPM型金属酶铜绿假单胞菌的检测分析

李进; 黎敏; 鲁卫平

doi:10.16016/j.1000-5404.202003323

您当前的位置：

首页 >

文章列表页 >

MALDI-TOF MS技术对VIM型和SPM型金属酶铜绿假单胞菌的检测分析

临床医学 | 更新时间：2026-01-09

- MALDI-TOF MS技术对VIM型和SPM型金属酶铜绿假单胞菌的检测分析
- Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry for detection of VIM- and SPM-type metallo-β-lactamase-producing Pseudomonas aeruginosa
- 第三军医大学学报 2020年42卷第16期页码：1663-1669
- 作者机构：
  
  400042 重庆, 陆军军医大学(第三军医大学)大坪医院检验科
- 作者简介：
  
  [ 鲁卫平, E-mail: luweiping19710416@163.com LU Weiping, E-mail: luweiping19710416@163.com
- 基金信息：
  
  陆军特色医学中心人才创新能力培养计划(2019CXJSC018)
- DOI：10.16016/j.1000-5404.202003323
  中图分类号： R372;R378.991;R446.5
- 收稿：2020-03-25，
  
  修回：2020-05-26，
  
  纸质出版：2020-08-30
- 稿件说明：
移动端阅览
李进, 黎敏, 鲁卫平. MALDI-TOF MS技术对VIM型和SPM型金属酶铜绿假单胞菌的检测分析[J]. 第三军医大学学报, 2020,42(16):1663-1669.

Jin LI, Min LI, Weiping LU. Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry for detection of VIM- and SPM-type metallo-β-lactamase-producing Pseudomonas aeruginosa[J]. Journal of Third Military Medical University, 2020, 42(16): 1663-1669.
李进, 黎敏, 鲁卫平. MALDI-TOF MS技术对VIM型和SPM型金属酶铜绿假单胞菌的检测分析[J]. 第三军医大学学报, 2020,42(16):1663-1669. DOI： 10.16016/j.1000-5404.202003323.

Jin LI, Min LI, Weiping LU. Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry for detection of VIM- and SPM-type metallo-β-lactamase-producing Pseudomonas aeruginosa[J]. Journal of Third Military Medical University, 2020, 42(16): 1663-1669. DOI： 10.16016/j.1000-5404.202003323.

摘要

目的

了解基质辅助激光解吸电离飞行时间质谱(matrix-assisted laser desorption/ionization time-of-flight mass spectrometry

MALDI-TOF MS)在检测金属酶铜绿假单胞菌(metallo-β-lactamase Pseudomonas aeruginosa

MBL-PA)类型中的作用。

方法

收集2015—2018年重庆某三甲医院临床分离的258株碳青霉烯类耐药铜绿假单胞菌进行EDTA协同试验和耐药基因检测

通过建立VIM型和SPM型MBL-PA识别模型数据库

对MALDI-TOF MS检测MBL-PA类型的灵敏度、特异度、阳性预测值和阴性预测值进行分析。

结果

258株碳青霉烯类耐药铜绿假单胞菌中EDTA表型初筛试验阳性106株

经PCR耐药基因检测得到VIM型40株

SPM型35株。通过MALDI-TOF MS识别模型检测出23/25 VIM型菌株

灵敏度为92.0%

特异性为100.0%

阳性预测值为100.0%

阴性预测值为93.7%。对于SPM型菌株

检测出16/20

灵敏度为80.0%

特异性为100.0%

阳性预测值均为100.0%

阴性预测值为88.2%。

结论

MALDI-TOF MS在检测MBL-PA方面比常规PCR技术更有优势

通过构建识别模型可快速准确地鉴别VIM型和SPM型MBL-PA

具备非常广的应用前景。

Abstract

Objective

To evaluate the performance of matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) for detecting VIM- and SPM-type metallo-β-lactamase-producing

Pseudomonas aeruginosa

(MBL-PA).

Methods

A total of 258 clinical isolates of carbapenem-resistant

P. aeruginosa

collected from our hospital from 2015 to 2018 were analyzed using EDTA test and PCR-based drug resistance gene detection. A VIM- and SPM-type recognition model database was established to assess the sensitivity

specificity

positive predictive value and negative predictive value of MALDI-TOF MS for detecting different types of MBL-PA.

Results

EDTA test detected 106 positive strains from the 258 clinical isolates of carbapenem-resistant

P. aeruginosa

and PCR-based drug resistance gene detection identified 40 VIM-type MBL-PA strains and 35 SPM-type MBL-PA strains. The MALDI-TOF MS recognition model detected 23 out of 25 VIM-type MBL-PA strains with a sensitivity of 92.0%

a specificity of 100.0%

a positive predictive value of 100.0%

and a negative predictive value of 93.7%

and identified 16 out of 20 SPM-type MBL-PA strains with a sensitivity of 80.0%

a specificity of 100.0%

a positive predictive value of 100.0%

and a negative predictive value of 88.2%.

Conclusion

MALDI-TOF MS has more advantages than conventional PCR in detecting the types of MBL-PA

and can quickly and accurately identify the VIM- and SPM-type MBL-PA after construction of a recognition model.

关键词

Keywords

references

LI J, HU W W, ZHANG F L, et al . Evaluation of matrix-assisted laser desorption/ionization time-of-flight mass spectrometry for identifying Burkholderia pseudomallei and Burkholderia thailandensis isolates[J ] . Eur J Clin Microbiol Infect Dis, 2019, 38(1): 191-196. DOI: 10.1007/s10096-018-3415-3.

GU Y F, LI Y, ZHANG X L, et al . A new method aimed to quickly identify pathogen and drug susceptibility test based on matrix-assisted laser desorption/ionization time of flight mass spectrometry combined with flow cytometry[J ] . Surg Infect (Larchmt), 2019, 20(3): 219-224. DOI: 10.1089/sur.2018.145.

HU Y Y, HUANG Y L, LIZOU Y Z, et al . Evaluation of Staphylococcus aureus subtyping module for methicillin-resistant Staphylococcus aureus detection based on matrix-assisted laser desorption ionization time-of-flight mass spectrometry[J ] . Front Microbiol, 2019, 10: 2504. DOI: 10.3389/fmicb.2019.02504.

RIBEIRO Á C D S, CROZATTI M T L, SILVA AA D, et al . Pseudomonas aeruginosa in the ICU: prevalence, resistance profile, and antimicrobial consumption[J ] . Rev Soc Bras Med Trop, 2019, 53: e20180498. DOI: 10.1590/0037-8682-0498-2018.

EL-MAHDY R, EL-KANNISHY G. Virulence factors ofcarbapenem-resistant Pseudomonas aeruginosa in hospital-acquired infections in mansoura, Egypt[J]. Infect Drug Resist, 2019, 12: 3455-3461. DOI: 10.2147/IDR.S222329.

李进, 胡韦维, 张峰领, 等.重庆某三甲医院2014-2016年铜绿假单胞菌耐药表型及金属酶、外膜孔蛋白耐药基因型分析[J].第三军医大学学报, 2017, 39(22): 2200-2205. DOI: 10.16016/j.1000-5404.201706013.

LI J, HU W W, ZHANG F L, et al . Drug resistance profiles and metallo-β-lactamase and outer membrane protein OprD2 genotypes of Pseudomonas aeruginosa: a 2014-2016 survey in a Grade 3 Class A general hospital in Chongqing[J ] . J Third Mil Med Univ, 2017, 39(22): 2200-2205. DOI: 10.16016/j.1000-5404.201706013.

曾婷, 曾凌, 曹先伟, 等.耐碳青霉烯类铜绿假单胞菌耐药基因与药敏分析[J].中华医院感染学杂志, 2019, 29(20): 3063-3067.

ZENG T, ZENG L, CAO X W, et al . Antimicrobial susceptibility and resistance genes of carbapenem-resistant Pseudomonas aeruginosa[J ] . Chin J Nosocomiol, 2019, 29(20): 3063-3067.

刘先林, 欧阳琳, 谢伟, 等.赣南地区耐碳青霉烯类药物铜绿假单胞菌耐药基因检测结果及其对药物的耐药性分析[J].抗感染药学, 2019, 16(9): 1512-1514.

LIU X L, OUYANG L, XIE W, et al . Detection results of resistance genes of carbapenem-resistant Pseudomonas aeruginosa and analysis of drug resistance in South Jiangxi Province. Anti-Infect Pharm, 2019, 16(9): 1512-1514.

骆远宁.耐碳青霉烯类铜绿假单胞菌的临床分布及其对抗菌药物的耐药性分析[J].临床合理用药杂志, 2019, 12(15): 93-94. DOI:10.15887/j.cnki.13-1389/r.2019.15.045.

LUO Y N. Clinical distribution ofcarbapenem-resistant Pseudomonas aeruginosa and its resistance to antibacterial drugs[J]. Chin J Clin Ration Drug Use, 2019, 12(15): 93-94. DOI: 10.15887/j.cnki.13-1389/r.2019.15.045.

许小敏, 范友芬, 张淳, 等.烧伤患者耐碳青霉烯类铜绿假单胞菌感染影响因素分析[J].中华医院感染学杂志, 2019, 29(12): 1791-1795.

XU X M, FAN Y F, ZHANG C, et al . Risk factors of carbapenem-resistant Pseudomonas aeruginosa infection in burn patients[J ] . Chin J Nosocomiol, 2019, 29(12): 1791-1795.

潘亚萍.外排泵MexAB-OprM在铜绿假单胞菌对碳青霉烯类抗生素耐药机制的研究[D].合肥: 安徽医科大学, 2017.

PAN Y P. Mechanism ofMexAB-OprM efflux pump in carbapenem-resistant Pseudomonas aeruginosa[D]. Hefei: Anhui Medical University, 2017.

袁莉莉, 丁百兴, 沈震, 等.耐碳青霉烯类铜绿假单胞菌的耐药性及分子流行病学研究[J].中国感染与化疗杂志, 2017, 17(3): 289-292. DOI: 10.16718/j.1009-7708.2017.03.012.

YUAN L L, DING B X, SHEN Z, et al . Antibiotic resistance and molecular epidemiology of carbapenem-resistant Pseudomonas aeruginosa[J ] . Chin J Infect Chemother, 2017, 17(3): 289-292. DOI: 10.16718/j.1009-7708.2017.03.012.

张智琪.氟喹诺酮药物诱导铜绿假单胞菌生物膜及耐药机制研究[D].太原: 山西医科大学, 2018.

ZHANG Z Q. Mechanism of resistance and biofilm of Pseudomonas aeruginosa induced by fluoroquinolones in vitro[D]. Taiyuan: Shanxi Medical University, 2018.

SONNABEND M S, KLEIN K, BEIER, et al . Identification of drug resistance determinants in a clinical isolate of Pseudomonas aeruginosa by high-density transposon mutagenesis[J ] . Antimicrob Agents Chemother, 2020, 64(3): e01771-e01719. DOI: 10.1128/AAC.01771-19.

徐雁峰, 王慧敏, 张冬.铜绿假单胞菌对碳青霉烯类抗生素耐药机制研究[J].包头医学院学报, 2019, 35(9): 108-112. DOI: 10.16833/j.cnki.jbmc.2019.09.048.

XU Y F, WANG H M, ZHANG D. Study on the resistance mechanism of Pseudomonas aeruginosa tocarbapenem antibiotics[J]. J Baotou Med Coll, 2019, 35(9): 108-112. DOI: 10.16833/j.cnki.jbmc.2019.09.048.

KHALILI Y, YEKANI M, GOLI H R, et al . Characterization of carbapenem-resistant but cephalosporin-susceptible Pseudomonas aeruginosa[J ] . Acta Microbiol Immunol Hung, 2019, 66(4): 529-540. DOI: 10.1556/030.66.2019.036.

PETROVA A, FEODOROVA Y, MITEVA-KATRANDZH-IEVA T, et al . First detected OXA-50 carbapenem-resistant clinical isolates Pseudomonas aeruginosa from Bulgaria and interplay between the expression of main efflux pumps, OprD and intrinsic AmpC[J ] . J Med Microbiol, 2019, 68(12): 1723-1731. DOI: 10.1099/jmm.0.001106.

ALCOLEA-MEDINA A, FERNANDEZ M T C, MONTIEL N, et al . An improved simple method for the identification of Mycobacteria by MALDI-TOF MS (Matrix-Assisted Laser Desorption-Ionization mass spectrometry)[J ] . Sci Rep, 2019, 9(1): 20216. DOI: 10.1038/s41598-019-56604-7.

PENG Y, ZHANG Q, XU C, et al . MALDI-TOF MS for the rapid identification and drug susceptibility testing of filamentous fungi[J ] . Exp Ther Med, 2019,18(6): 4865-4873. DOI: 10.3892/etm.2019.8118.

SAMARANAYAKE W A M P, DEMPSEY S, HOWARD-JONES A R, et al . Rapid d irect identification of positive paediatric blood cultures by MALDI-TOF MS technology and its clinical impact in the paediatric hospital setting[J ] . BMC Res Notes, 2020, 13(1): 12. DOI: 10.1186/s13104-019-4861-4.

PIAMSOMBOON P, JARESITTHIKUNCHAI J, HUNG T Q, et al . Identification of bacterial pathogens in cultured fish with a custom peptide database constructed by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS)[J ] . BMC Vet Res, 2020, 16(1): 52. DOI: 10.1186/s12917-020-2274-1.

QUÉRO L, COURAULT P, CELLIÈRE B, et al . Application of MALDI-TOF MS to species complex differentiation and strain typing of food related fungi: Case studies with Aspergillus section Flavi species and Penicillium roqueforti isolates[J ] . Food Microbiol, 2020, 86: 103311. DOI: 10.1016/j.fm.2019.103311.

LUO LL, LIU W J, LI B, et al . Evaluation of matrix-assisted laser desorption ionization-time of flight mass spectrometry for identification of Mycobacterium abscessus subspecies according to whole-genome sequencing[J ] . J Clin Microbiol, 2016, 54(12): 2982-2989. DOI: 10.1128/JCM.01151-16.

YOO G Y, KIM E, KANG H, et al . Mass spectrometric investigation of concentration-dependent effect of curcumin and oxidative stress on intracellular glutathione levels[J ] . Anal Bioanal Chem, 2020, 412(12): 2873-2880. DOI: 10.1007/s00216-020-02524-9.

SÁNCHEZ-JUANES F, TEIXEIRA-MARTÍN V, GONZÁLEZ-BUITRAGO J M, et al . Identification of species and subspecies of lactic acid bacteria present in Spanish cheeses type "torta" by MALDI-TOF MS and pheS gene analyses[J ] . Microorganisms, 2020, 8(2): E301. DOI: 10.3390/microorganisms8020301.

GOU JJ, LIU N, GUO L H, et al . Carbapenem-resistant Enterobacter hormaechei ST1103 with IMP-26 carbapenemase and ESBL gene blaSHV-178[J ] . Infect Drug Resist, 2020, 13: 597-605. DOI: 10.2147/IDR.S232514.

浏览量

下载量

CSCD

文章被引用时，请邮件提醒。

提交

工具集

关联资源

暂无数据