细菌超广谱β内酰胺酶检测方法的研究
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摘要
超广谱β-内酰胺酶(ESBLs)菌株引起的耐药率逐渐增高,本文主要从定性和定量两个方面概述了ESBLs检测方法的原理和步骤。其中,定性的方法包括药敏纸片法、MIC法、自动仪器法、比色法、质谱法以及基因检测法等;定量即利用超声波,破碎提取细菌体内的超广谱β-内酰胺酶,对其进行活性测定,计算确切的活性单位。并对各种方法的操作过程、检测灵敏性、检测速度以及检测准确性等方面进行比较,以期为产ESBLs细菌感染的监测和控制提供参考。
Antimicrobial resistance caused by Extended-spectrum β-lactamases(ESBLs)is on the rise. This paper summarizes principle and procedure of qualitative and quantitative detection methods for ESBLs.Qualitative methods include drug sensitive test,MIC method,automatic system,colorimetry,spectrometry and gene detection,etc.Quantitative procedure is to determine the biological activity of the Extended-spectrum β-lactamases(ESBLs)extracted by ultrasonication,and calculate the exact enzymatic activity of ESBLs.And all methods were compared on operation mode,sensitivity of detection,inspection speed and the accuracy of test,expecting to provide references for monitoring and control of ESBLs-producing bacterial infection.
Antimicrobial resistance caused by Extended-spectrum β-lactamases(ESBLs)is on the rise. This paper summarizes principle and procedure of qualitative and quantitative detection methods for ESBLs.Qualitative methods include drug sensitive test,MIC method,automatic system,colorimetry,spectrometry and gene detection,etc.Quantitative procedure is to determine the biological activity of the Extended-spectrum β-lactamases(ESBLs)extracted by ultrasonication,and calculate the exact enzymatic activity of ESBLs.And all methods were compared on operation mode,sensitivity of detection,inspection speed and the accuracy of test,expecting to provide references for monitoring and control of ESBLs-producing bacterial infection.
引文
[1]BRADFORD,P A.Extended-Spectrum β-Lactamases in the 21st Century:Characterization,Epidemiology,and Detection of This Important Resistance Threat[J].Clinical Microbiology Reviews,2001,14(4):933-951.
[2]陈民钧.美国临床实验室标准化委员会2004年版有关药敏试验标准化更新要点[J].中华检验医学杂志,2004,27(9):608-610.
[3]孔庆莲,于秀娟,姚苹.产超广谱β-内酰胺酶大肠埃希菌和肺炎克雷伯菌的检测[J].中华检验医学杂志,2000,23(6):360-363.
[4]史跃杰,杨玉林.3-氨基苯硼酸改良方法检测超广谱β-内酰胺酶[J].中国卫生检验杂志,2008,18(7):1271-1273.
[5]BEDENIC B,RANDEGGER C,BORAS A,et al.Comparison of five different methods for detection of SHV extended-spectrum beta-lactamases[J].Journal of Chemotherapy,2014,13(1):24-33.
[6]KASSIM A,OMUSE G,PREMJI Z,et al.Comparison of Clinical Laboratory Standards Institute and European Committee on Antimicrobial Susceptibility Testing guidelines for the interpretation of antibiotic susceptibility at a University teaching hospital in Nairobi,Kenya:a cross-sectional study[J].Annals of Clinical Microbiology and Antimicrobials,2016,15(1):21.
[7]杜军,李敏,崔景荣,等.4种检测超广谱β-内酰胺酶方法的比较[J].宁夏大学学报(自然科学版),2009,30(3):268-270.
[8]施菁玲,李从荣,李红霞,等.产超广谱β内酰胺酶细菌的自动化检测及耐药性研究[J].中国人兽共患病学报,2000,16(4):76-77.
[9]李虹玲,刘文恩.SHV型超广谱β-内酰胺酶的研究进展[J].实用预防医学,2007,14(1):253-255.
[10]董佳琳,施立煌.ATBG-5肠杆菌药敏条对超广谱β-内酰胺酶的检测分析[J].临床医学工程,2012,19(11):1872-1873.
[11]肖于飞,张嵘,徐志江,等.Vitek1、Vitek2和BD Phoenix系统对超广谱β-内酰胺酶的检测和评价[J].临床检验杂志,2006,24(4):250-252.
[12]张军民,罗燕萍,崔岩,等.临床评价VITEK2高级专家系统对常见细菌β内酰胺耐药表型的检测和分析[J].中华检验医学杂志,2003,26(4):233-235.
[13]WASSEF M A,RAHMAN E M A.Chromogenic Cica-βTesting for Detection of Extended-Spectrum and AmpCβ-Lactamases Among Cefoxitin-Resistant Isolates[J].Laboratory Medicine,2013,44(1):25-28.
[14]RENVOISE A,DECRE D,AMARSY-GUERLE R,et al.Evaluation of theβLacta Test,a Rapid Test Detecting Resistance to Third-Generation Cephalosporins in Clinical Strains of Enterobacteriaceae[J].Journal of Clinical Microbiology,2013,51(12):4012-4017.
[15]NORDMANN P D L P L.Rapid Detection of Extended-Spectrum-β-Lactamase-Producing Enterobacteriaceae[J].Journal of Clinical Microbiology,2012,50(9):3016-3022.
[16]LI B,GUO T,QU F,et al.Matrix-assisted laser desorption ionization:time of flight mass spectrometry-identified models for detection of ESBL-producing bacterial strains[J].Med Sci Monit Basic Res,2014(20):176-183.
[17]SPARBIER K,SCHUBERT S,WELLER U,et al.Matrix-assisted laser desorption ionization-time of flight mass spectrometry-based functional assay for rapid detection of resistance againstβ-lactam antibiotics[J].Journal of Clinical Microbiology,2012,50(3):927.
[18]JENI V,JEAN-MARC C,WHALEY M J,et al.Development of Real-Time PCR Methods for the Detection of Bacterial Meningitis Pathogens without DNA Extraction[J].PLOS ONE,2016,11(2):e0147765.
[19]SOUVEREIN D,EUSER S M,VAN DER REIJDEN W A,et al.Clinical sensitivity and specificity of the Check-Points Check-Direct ESBL Screen for BD MAX,a real-time PCR for direct ESBL detection from rectal swabs[J].Journal of Antimicrobial Chemotherapy,2017,72(9):2512-2518.
[20]CARD R,ZHANG J,DAS P,et al.Evaluation of an Expanded Microarray for Detecting Antibiotic Resistance Genes in a Broad Range of Gram-Negative Bacterial Pathogens[J].Antimicrobial Agents and Chemotherapy,2013,57(1):458-465.
[21]MANCINI N,INFURNARI L,GHIDOLI N,et al.Potential Impact of a Microarray-Based Nucleic Acid Assay for Rapid Detection of Gram-Negative Bacteria and Resistance Markers in Positive Blood Cultures[J].Journal of Clinical Microbiology,2014,52(4):1242-1245.
[22]朱卫民,钱元恕.TEM型β-内酰胺酶动力学研究[J].中国抗生素杂志,2001,26(6):460-463.
[23]RICHMOND M H.The beta-lactamase stability of a novel beta-lactam antibiotic containing a 7 alpha-methoxyoxacephem nucleus[J].Journal of Antimicrobial Chemotherapy,1980,6(4):445-453.
[24]朱卫民,陈勇川,钱元恕.肺炎克雷伯氏菌AmpCβ-内酰胺酶动力学及抑酶效应的研究[J].中国抗生素杂志,2003,28(6):351-353、372.
[25]CORMICAN M G,MARSHALL S A,JONES R N.Detection of extended-spectrum beta-lactamase(ESBL)-producing strains by the Etest ESBL screen[J].Journal of Clinical Microbiology,1996,34(8):1880-1884.
[2]陈民钧.美国临床实验室标准化委员会2004年版有关药敏试验标准化更新要点[J].中华检验医学杂志,2004,27(9):608-610.
[3]孔庆莲,于秀娟,姚苹.产超广谱β-内酰胺酶大肠埃希菌和肺炎克雷伯菌的检测[J].中华检验医学杂志,2000,23(6):360-363.
[4]史跃杰,杨玉林.3-氨基苯硼酸改良方法检测超广谱β-内酰胺酶[J].中国卫生检验杂志,2008,18(7):1271-1273.
[5]BEDENIC B,RANDEGGER C,BORAS A,et al.Comparison of five different methods for detection of SHV extended-spectrum beta-lactamases[J].Journal of Chemotherapy,2014,13(1):24-33.
[6]KASSIM A,OMUSE G,PREMJI Z,et al.Comparison of Clinical Laboratory Standards Institute and European Committee on Antimicrobial Susceptibility Testing guidelines for the interpretation of antibiotic susceptibility at a University teaching hospital in Nairobi,Kenya:a cross-sectional study[J].Annals of Clinical Microbiology and Antimicrobials,2016,15(1):21.
[7]杜军,李敏,崔景荣,等.4种检测超广谱β-内酰胺酶方法的比较[J].宁夏大学学报(自然科学版),2009,30(3):268-270.
[8]施菁玲,李从荣,李红霞,等.产超广谱β内酰胺酶细菌的自动化检测及耐药性研究[J].中国人兽共患病学报,2000,16(4):76-77.
[9]李虹玲,刘文恩.SHV型超广谱β-内酰胺酶的研究进展[J].实用预防医学,2007,14(1):253-255.
[10]董佳琳,施立煌.ATBG-5肠杆菌药敏条对超广谱β-内酰胺酶的检测分析[J].临床医学工程,2012,19(11):1872-1873.
[11]肖于飞,张嵘,徐志江,等.Vitek1、Vitek2和BD Phoenix系统对超广谱β-内酰胺酶的检测和评价[J].临床检验杂志,2006,24(4):250-252.
[12]张军民,罗燕萍,崔岩,等.临床评价VITEK2高级专家系统对常见细菌β内酰胺耐药表型的检测和分析[J].中华检验医学杂志,2003,26(4):233-235.
[13]WASSEF M A,RAHMAN E M A.Chromogenic Cica-βTesting for Detection of Extended-Spectrum and AmpCβ-Lactamases Among Cefoxitin-Resistant Isolates[J].Laboratory Medicine,2013,44(1):25-28.
[14]RENVOISE A,DECRE D,AMARSY-GUERLE R,et al.Evaluation of theβLacta Test,a Rapid Test Detecting Resistance to Third-Generation Cephalosporins in Clinical Strains of Enterobacteriaceae[J].Journal of Clinical Microbiology,2013,51(12):4012-4017.
[15]NORDMANN P D L P L.Rapid Detection of Extended-Spectrum-β-Lactamase-Producing Enterobacteriaceae[J].Journal of Clinical Microbiology,2012,50(9):3016-3022.
[16]LI B,GUO T,QU F,et al.Matrix-assisted laser desorption ionization:time of flight mass spectrometry-identified models for detection of ESBL-producing bacterial strains[J].Med Sci Monit Basic Res,2014(20):176-183.
[17]SPARBIER K,SCHUBERT S,WELLER U,et al.Matrix-assisted laser desorption ionization-time of flight mass spectrometry-based functional assay for rapid detection of resistance againstβ-lactam antibiotics[J].Journal of Clinical Microbiology,2012,50(3):927.
[18]JENI V,JEAN-MARC C,WHALEY M J,et al.Development of Real-Time PCR Methods for the Detection of Bacterial Meningitis Pathogens without DNA Extraction[J].PLOS ONE,2016,11(2):e0147765.
[19]SOUVEREIN D,EUSER S M,VAN DER REIJDEN W A,et al.Clinical sensitivity and specificity of the Check-Points Check-Direct ESBL Screen for BD MAX,a real-time PCR for direct ESBL detection from rectal swabs[J].Journal of Antimicrobial Chemotherapy,2017,72(9):2512-2518.
[20]CARD R,ZHANG J,DAS P,et al.Evaluation of an Expanded Microarray for Detecting Antibiotic Resistance Genes in a Broad Range of Gram-Negative Bacterial Pathogens[J].Antimicrobial Agents and Chemotherapy,2013,57(1):458-465.
[21]MANCINI N,INFURNARI L,GHIDOLI N,et al.Potential Impact of a Microarray-Based Nucleic Acid Assay for Rapid Detection of Gram-Negative Bacteria and Resistance Markers in Positive Blood Cultures[J].Journal of Clinical Microbiology,2014,52(4):1242-1245.
[22]朱卫民,钱元恕.TEM型β-内酰胺酶动力学研究[J].中国抗生素杂志,2001,26(6):460-463.
[23]RICHMOND M H.The beta-lactamase stability of a novel beta-lactam antibiotic containing a 7 alpha-methoxyoxacephem nucleus[J].Journal of Antimicrobial Chemotherapy,1980,6(4):445-453.
[24]朱卫民,陈勇川,钱元恕.肺炎克雷伯氏菌AmpCβ-内酰胺酶动力学及抑酶效应的研究[J].中国抗生素杂志,2003,28(6):351-353、372.
[25]CORMICAN M G,MARSHALL S A,JONES R N.Detection of extended-spectrum beta-lactamase(ESBL)-producing strains by the Etest ESBL screen[J].Journal of Clinical Microbiology,1996,34(8):1880-1884.