Biotechnology Bulletin ›› 2022, Vol. 38 ›› Issue (9): 59-71.doi: 10.13560/j.cnki.biotech.bull.1985.2022-0854
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ZHAO Yan-kun1(), LIU Hui-min2, MENG Lu2, WANG Cheng1, WANG Jia-qi2, ZHENG Nan2()
Received:
2022-07-10
Online:
2022-09-26
Published:
2022-10-11
Contact:
ZHENG Nan
E-mail:yankunzhao90@163.com;zhengnan@caas.cn
ZHAO Yan-kun, LIU Hui-min, MENG Lu, WANG Cheng, WANG Jia-qi, ZHENG Nan. Research Progress in Heteroresistance of Escherichia coli[J]. Biotechnology Bulletin, 2022, 38(9): 59-71.
方法 Methods | 优点 Advantages | 缺点 Disadvantages | 参考文献 Reference |
---|---|---|---|
E-test法、K-B纸片扩散法 E-test and Kirby-Bauer test | (1)传统、经典的药敏试验方法; (2)可用于异质性耐药的初筛; (3)操作简便、成本较低、结果直观易判读。 | (1)由于平板上的细胞密度低,不易检出低频率出现的耐药亚群; (2)可信度低,假阳性、假阴性检出率较高。 | [ |
M-E-test法M-E-test method | (1)可用于异质性耐药的大规模筛选; (2)操作简单。 | 培养板上细胞密度越高,检测异质性耐药亚群的概率越高,但检测的准确率不稳定,易造成误判。 | [ |
GRD E-test、琼脂筛选 GRD E-test and agar screen | (1)临床实验室常规使用的药敏试验方法; (2)用于观察某种程度的耐药性和异质性耐药性之间存在强烈相关性的简单筛选方法(hVISA)。 | (1)无法检测异质性耐药; (2)缺乏有关亚群频率和MIC的信息。 | [ |
微量肉汤稀释法 Broth microdilution(BMD) | 临床实验室常规使用的药敏试验方法。 | (1)在存在耐药亚群的情况下,MIC的读数高度依赖于耐药细菌的接种密度和生长速率; (2)没有关于耐药亚群频率的信息,其MIC信息较差; (3)操作繁琐。 | [ |
琼脂稀释 Agar dilution | 临床实验室常规使用的药敏试验方法。 | (1)使用耐药细菌的单个小菌落有可能错过低频率的亚群; (2)如果检测到异质性耐药,则提供有关耐药亚群的频率和MIC的少许信息; (3)耗时、费力。 | [ |
群体谱分析法 PAP test | (1)检测异质性耐药较可靠的方法;(2)能够提供有关耐药亚群频率和MIC的有效信息; (3)可定量分析、准确度高、稳定性好、可重复性好。 | (1)操作繁琐且成本昂贵; (2)耗时耗力; (3)技术性强,临床推广应用困难。 | [ |
微量稀释群体谱分析法 Microdilution PAP test | (1)比常规PAP测试更快、更便宜; (2)可得知耐药亚群的频率和MIC。 | 检测的效果取决于细胞密度,误差较大。 | [ |
改良群体谱分析曲线法 PAP-AUC | (1)hVISA检测的黄金标准; (2)准确性高。 | (1)步骤繁琐且成本昂贵; (2)参考hVISA菌株可能会对不同药物浓度具有高度依赖性的耐药亚群分离株进行错误分类,造成假阳性或假阴性结果。 | [ |
自动肉汤系统(如VITEK 2、MicroScan、WalkAway) Automated broth systems(for example,VITEK 2,MicroScan,and WalkAway) | 临床实验室常规使用药敏试验方法。 | 在检测耐药亚群方面效果不佳。 | [ |
分子检测方法(如探针分析和GeneXpert-Xpert-MTB/RIF-Ultra) Molecular detection methods(for example,line probe assays,GeneXpert Xpert MTB/RIF Ultra) | (1)与传统或自动检测方法相比,更能快速检测异质性耐药; (2)能够准确检测细菌亚群的点突变。 (3)有时可直接用于患者样本(如,结核分枝杆菌的痰样本)。 | (1)并非所有探针分析系统都能检测耐药亚群,检测效率和稳定性不稳定; (2)依赖于对耐药基因的全面了解,会造成结果差异较大,不适用于所有细菌。 | [ |
Table 1 Comparison of detection methods for heteroresistance
方法 Methods | 优点 Advantages | 缺点 Disadvantages | 参考文献 Reference |
---|---|---|---|
E-test法、K-B纸片扩散法 E-test and Kirby-Bauer test | (1)传统、经典的药敏试验方法; (2)可用于异质性耐药的初筛; (3)操作简便、成本较低、结果直观易判读。 | (1)由于平板上的细胞密度低,不易检出低频率出现的耐药亚群; (2)可信度低,假阳性、假阴性检出率较高。 | [ |
M-E-test法M-E-test method | (1)可用于异质性耐药的大规模筛选; (2)操作简单。 | 培养板上细胞密度越高,检测异质性耐药亚群的概率越高,但检测的准确率不稳定,易造成误判。 | [ |
GRD E-test、琼脂筛选 GRD E-test and agar screen | (1)临床实验室常规使用的药敏试验方法; (2)用于观察某种程度的耐药性和异质性耐药性之间存在强烈相关性的简单筛选方法(hVISA)。 | (1)无法检测异质性耐药; (2)缺乏有关亚群频率和MIC的信息。 | [ |
微量肉汤稀释法 Broth microdilution(BMD) | 临床实验室常规使用的药敏试验方法。 | (1)在存在耐药亚群的情况下,MIC的读数高度依赖于耐药细菌的接种密度和生长速率; (2)没有关于耐药亚群频率的信息,其MIC信息较差; (3)操作繁琐。 | [ |
琼脂稀释 Agar dilution | 临床实验室常规使用的药敏试验方法。 | (1)使用耐药细菌的单个小菌落有可能错过低频率的亚群; (2)如果检测到异质性耐药,则提供有关耐药亚群的频率和MIC的少许信息; (3)耗时、费力。 | [ |
群体谱分析法 PAP test | (1)检测异质性耐药较可靠的方法;(2)能够提供有关耐药亚群频率和MIC的有效信息; (3)可定量分析、准确度高、稳定性好、可重复性好。 | (1)操作繁琐且成本昂贵; (2)耗时耗力; (3)技术性强,临床推广应用困难。 | [ |
微量稀释群体谱分析法 Microdilution PAP test | (1)比常规PAP测试更快、更便宜; (2)可得知耐药亚群的频率和MIC。 | 检测的效果取决于细胞密度,误差较大。 | [ |
改良群体谱分析曲线法 PAP-AUC | (1)hVISA检测的黄金标准; (2)准确性高。 | (1)步骤繁琐且成本昂贵; (2)参考hVISA菌株可能会对不同药物浓度具有高度依赖性的耐药亚群分离株进行错误分类,造成假阳性或假阴性结果。 | [ |
自动肉汤系统(如VITEK 2、MicroScan、WalkAway) Automated broth systems(for example,VITEK 2,MicroScan,and WalkAway) | 临床实验室常规使用药敏试验方法。 | 在检测耐药亚群方面效果不佳。 | [ |
分子检测方法(如探针分析和GeneXpert-Xpert-MTB/RIF-Ultra) Molecular detection methods(for example,line probe assays,GeneXpert Xpert MTB/RIF Ultra) | (1)与传统或自动检测方法相比,更能快速检测异质性耐药; (2)能够准确检测细菌亚群的点突变。 (3)有时可直接用于患者样本(如,结核分枝杆菌的痰样本)。 | (1)并非所有探针分析系统都能检测耐药亚群,检测效率和稳定性不稳定; (2)依赖于对耐药基因的全面了解,会造成结果差异较大,不适用于所有细菌。 | [ |
抗菌药物 名称 Antibiotic | 样品(来源) Sample (Sources) | 样本数 Number of samples | 异质性耐药测定方法Method for determination of heteroresistance | 异质性耐药率 Prevalence of heteroresistance/% | 耐药率Prevalence of resistance/% | 异质性耐药亚群的频次 Frequency of heteroresistant subpopulations | 国家/地区Country /Region | 文献 Reference |
---|---|---|---|---|---|---|---|---|
亚胺培南 Imipenem | 医院分离株 | 11 | E-test(抑菌圈内菌落生长初步判定异质性) PAP(异质性耐药以MIC/NIC≥8来确定) | 0 | 0 | ND | 瑞典 | [ |
厄他培南 Ertapenem | 11 | 27.3 | 0 | 4.20×10-7 - 1.10×10-5 | ||||
美罗培南 Meropenem | 11 | 0 | 0 | ND | ||||
多利培南 Doripenem | 11 | 0 | 0 | ND | ||||
厄他培南 Ertapenem | 医院分离株 | 140 | 改良PAP | 35 | 2 | ND | 美国 | [ |
亚胺培南 Imipenem | 20 | 25 | ||||||
美罗培南 Meropenem | 100 | 30 | ||||||
亚胺培南 Imipenem | 医院分离株 | 332 | 纸片扩散,E-test(抑菌圈内菌落生长确定为异质性) | 25 | 0 | 8.00×10-8- 1.80×10-7 | 中国重庆 | [ |
厄他培南 Ertapenem | 17.2 | 0.6 | ||||||
美罗培南 Meropenem | 3.9 | |||||||
亚胺培南 Imipenem | 医院分离株 | 200 | E-test(抑菌圈内菌落生长确定为异质性) PAP(异质性耐药以MIC/NIC≥8来确定) | 29 | 0 | ND | 中国广西 | [ |
厄他培南 Ertapenem | 20.5 | |||||||
美罗培南 Meropenem | 6 | 0.5 | ||||||
亚胺培南 Imipenem | 血液、尿液和痰液 | 850 | K-B纸片法 E-test PAP(异质性耐药以MIC/NIC≥8来确定) | 20.5 | 0 | ND | 中国广东 | [ |
厄他培南 Ertapenem | 17.5 | 1.5 | ||||||
美罗培南 Meropenem | 7 | 0.5 |
Table 2 Prevalence of heteroresistance of E. coli to carbapenems
抗菌药物 名称 Antibiotic | 样品(来源) Sample (Sources) | 样本数 Number of samples | 异质性耐药测定方法Method for determination of heteroresistance | 异质性耐药率 Prevalence of heteroresistance/% | 耐药率Prevalence of resistance/% | 异质性耐药亚群的频次 Frequency of heteroresistant subpopulations | 国家/地区Country /Region | 文献 Reference |
---|---|---|---|---|---|---|---|---|
亚胺培南 Imipenem | 医院分离株 | 11 | E-test(抑菌圈内菌落生长初步判定异质性) PAP(异质性耐药以MIC/NIC≥8来确定) | 0 | 0 | ND | 瑞典 | [ |
厄他培南 Ertapenem | 11 | 27.3 | 0 | 4.20×10-7 - 1.10×10-5 | ||||
美罗培南 Meropenem | 11 | 0 | 0 | ND | ||||
多利培南 Doripenem | 11 | 0 | 0 | ND | ||||
厄他培南 Ertapenem | 医院分离株 | 140 | 改良PAP | 35 | 2 | ND | 美国 | [ |
亚胺培南 Imipenem | 20 | 25 | ||||||
美罗培南 Meropenem | 100 | 30 | ||||||
亚胺培南 Imipenem | 医院分离株 | 332 | 纸片扩散,E-test(抑菌圈内菌落生长确定为异质性) | 25 | 0 | 8.00×10-8- 1.80×10-7 | 中国重庆 | [ |
厄他培南 Ertapenem | 17.2 | 0.6 | ||||||
美罗培南 Meropenem | 3.9 | |||||||
亚胺培南 Imipenem | 医院分离株 | 200 | E-test(抑菌圈内菌落生长确定为异质性) PAP(异质性耐药以MIC/NIC≥8来确定) | 29 | 0 | ND | 中国广西 | [ |
厄他培南 Ertapenem | 20.5 | |||||||
美罗培南 Meropenem | 6 | 0.5 | ||||||
亚胺培南 Imipenem | 血液、尿液和痰液 | 850 | K-B纸片法 E-test PAP(异质性耐药以MIC/NIC≥8来确定) | 20.5 | 0 | ND | 中国广东 | [ |
厄他培南 Ertapenem | 17.5 | 1.5 | ||||||
美罗培南 Meropenem | 7 | 0.5 |
抗菌药物 名称 Antibiotic | 样品(来源) Sample(Sources) | 样本数 Number of samples | 异质性耐药测定方法 Method for determination of heteroresistance | 异质性耐药率 Prevalence of heteroresistance/% | 耐药率 Prevalence of resistance/% | 异质性耐药亚群的频次 Frequency of heteroresistant subpopulations | 国家/地区Country / Region | 文献 Reference |
---|---|---|---|---|---|---|---|---|
黏菌素 Colistin | 临床分离株 | 11 | E-test(抑菌圈内菌落生长初步判定异质性) PAP(异质性耐药以MIC/NIC≥8来确定) | 0 | 0 | ND | 瑞典 | [ |
多黏菌素B Polymyxin B | 11 | 0 | 0 | ND | ||||
黏菌素 Colistin | 血液分离株 | 292 | 琼脂筛选[菌落生长在黏菌素(4 mg/L)确定为异质性耐药)] | 3.4 | 0.3 | 1.25×10-8-1.10×10-5 | 匈牙利 | [ |
黏菌素 Colistin | 医院分离株 | 291 | PAP(异质性耐药以MIC/NIC≥8来确定) PAP | 1.37 | 0.69 | 4.00×10-7-4.00×10-6 | 中国温州 | [ |
黏菌素 Colistin | 猪病料样本、猪肛门拭子 | 177 | PAP(异质性耐药以MIC/NIC≥4来确定) | 91.67 | 0 | 6.61×10-7-2.57×10-6 | 中国郑州 | [ |
Table 3 Prevalence of heteroresistance of E. coli to colistin antibiotics
抗菌药物 名称 Antibiotic | 样品(来源) Sample(Sources) | 样本数 Number of samples | 异质性耐药测定方法 Method for determination of heteroresistance | 异质性耐药率 Prevalence of heteroresistance/% | 耐药率 Prevalence of resistance/% | 异质性耐药亚群的频次 Frequency of heteroresistant subpopulations | 国家/地区Country / Region | 文献 Reference |
---|---|---|---|---|---|---|---|---|
黏菌素 Colistin | 临床分离株 | 11 | E-test(抑菌圈内菌落生长初步判定异质性) PAP(异质性耐药以MIC/NIC≥8来确定) | 0 | 0 | ND | 瑞典 | [ |
多黏菌素B Polymyxin B | 11 | 0 | 0 | ND | ||||
黏菌素 Colistin | 血液分离株 | 292 | 琼脂筛选[菌落生长在黏菌素(4 mg/L)确定为异质性耐药)] | 3.4 | 0.3 | 1.25×10-8-1.10×10-5 | 匈牙利 | [ |
黏菌素 Colistin | 医院分离株 | 291 | PAP(异质性耐药以MIC/NIC≥8来确定) PAP | 1.37 | 0.69 | 4.00×10-7-4.00×10-6 | 中国温州 | [ |
黏菌素 Colistin | 猪病料样本、猪肛门拭子 | 177 | PAP(异质性耐药以MIC/NIC≥4来确定) | 91.67 | 0 | 6.61×10-7-2.57×10-6 | 中国郑州 | [ |
Fig. 1 Recommended scheme for determination of heteroresistance(Modified from reference[31]) (1)Disc diffusion assays should be performed according to standardized procedures for antimicrobial susceptibility testing as recommended by agencies such as CLSI and BSAC.(2)PAP by CFU counts should be performed by plating aliquots of 10-fold serially diluted bacterial cultures on antibiotic-containing agar plates.(3)Agar plate preparation should follow standardized guidelines for MIC determination by agar dilution assays. Turbidimetric PAP should follow standard guidelines for MIC determination by the broth dilution technique,with the exception of turbidimetric quantification of bacterial growth at each antibiotic concentration
抗菌药物 Antibiotic | 异质性耐药机制 Heteroresistance mechanism | 参考文献 References |
---|---|---|
头孢菌素类 Cephalosporins | 在携带绿色荧光蛋白基因的质粒上将编码头孢菌素水解酶的blaCTX-M-14基因导入敏感的E.coli菌株,由于与绿色荧光蛋白的融合,在单细胞水平上出现了异质性耐药。 | [ |
四环素类 Tetracyclines | 耐药亚群中外排泵相关基因如AcrAB-Tolc、OqxAB、acrAB、oqxAB等表达水平增加或活性增强有关。 | [ |
磷霉素 Fosfomycin | 耐药亚群中转运调控基因突变导致磷霉素摄取系统缺陷及磷霉素耐药基因murA过度表达是引起异质性耐药的原因 | [ |
Table 4 Heteroresistance mechanism of E.coli to other antibiotics
抗菌药物 Antibiotic | 异质性耐药机制 Heteroresistance mechanism | 参考文献 References |
---|---|---|
头孢菌素类 Cephalosporins | 在携带绿色荧光蛋白基因的质粒上将编码头孢菌素水解酶的blaCTX-M-14基因导入敏感的E.coli菌株,由于与绿色荧光蛋白的融合,在单细胞水平上出现了异质性耐药。 | [ |
四环素类 Tetracyclines | 耐药亚群中外排泵相关基因如AcrAB-Tolc、OqxAB、acrAB、oqxAB等表达水平增加或活性增强有关。 | [ |
磷霉素 Fosfomycin | 耐药亚群中转运调控基因突变导致磷霉素摄取系统缺陷及磷霉素耐药基因murA过度表达是引起异质性耐药的原因 | [ |
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