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Detection of various beta-Lactamases in Escherichia coli and Klebsiella sp.: A study from Tertiary Care Centre of North India
Indian Journal of Medical Microbiology ( IF 1.4 ) Pub Date : 2020-07-01 , DOI: 10.4103/ijmm.ijmm_20_253 Varsha Gupta 1 , Meenakshi Singh 1 , Priya Datta 2 , Anku Goel 1 , Sanjay Singh 3 , Kashinath Prasad 3 , Jagdish Chander 1
Objective: The emergence of carbapenem-resistant Escherichia coli and Klebsiella species is a global threat. We aimed to compare two phenotypic methods and evaluate the genotypic method for the detection of beta-lactamases produced by E. coli and Klebsiella spp. Materials and Methods: One hundred and twenty-six E. coli and Klebsiella isolates were examined for phenotypic production of beta-lactamases by using disc diffusion, combined disc test (CDT) and modified carbapenem inactivation method (mCIM). All strains were also studied for the presence of various genes by polymerase chain reaction. Results: Out of 126 isolates, 96% of the isolates were extended-spectrum β-lactamase (ESBL) producers based on the presence of various ESBL genes. CDT method showed higher number of total (89%) carbapenemases in comparison to mCIM (81%). Among carbapenemases none of the isolates were Klebsiella pneumoniae carbapenemase producer by CDT, while 69% isolates were metallo-beta-lactamase (MBL) producers. Another method, mCIM/ethylene diamine tetraacetic acid mCIM showed 100% agreement for MBL detection. As regards, AmpC and class D carbapenemases; 0.04% and 16% positivity was detected, respectively, based on CDT method. Molecular analysis revealed 91% of the isolates harbouring carbapenemase genes. blaNDMwas the most common gene detected followed byblaOXA-48. Nine of the blaNDM-positive isolates also possessed blaOXA-48gene. Conclusion: Our finding shows high percentages of ESBL and carbapenemases in E. coli and Klebsiella spp. Among phenotypic methods, CDT seems to be a better choice as prevalence of carbapenemases shows lots of variation in our country. For Class B enzymes, both CDT and mCIM/eCIM can be used in the routine laboratories.
中文翻译：
大肠杆菌和克雷伯氏菌中各种 β-内酰胺酶的检测：北印度三级护理中心的一项研究
目的：耐碳青霉烯类大肠杆菌和克雷伯氏菌的出现对全球构成威胁。我们的目的是比较两种表型方法并评估用于检测大肠杆菌和克雷伯菌属产生的 β-内酰胺酶的基因型方法。材料和方法：通过使用纸片扩散、组合纸片测试 (CDT) 和改良碳青霉烯灭活方法 (mCIM)，检查 126 株大肠杆菌和克雷伯氏菌分离株的 β-内酰胺酶表型产生情况。还通过聚合酶链式反应研究了所有菌株中各种基因的存在。结果：在 126 个分离株中，基于各种 ESBL 基因的存在，96% 的分离株是超广谱 β-内酰胺酶 (ESBL) 生产者。 CDT 方法显示碳青霉烯酶总数 (89%) 高于 mCIM (81%)。根据 CDT，在碳青霉烯酶中，没有分离株是肺炎克雷伯菌碳青霉烯酶生产者，而 69% 的分离株是金属-β-内酰胺酶 (MBL) 生产者。另一种方法，mCIM/乙二胺四乙酸 mCIM 对 MBL 检测显示 100% 一致性。至于AmpC和D类碳青霉烯酶；基于CDT方法，分别检测到0.04%和16%的阳性率。分子分析显示 91% 的分离株携带碳青霉烯酶基因。 bla NDM是检测到的最常见基因，其次是bla OXA-48 。 9 个bla NDM阳性分离株也具有bla OXA-48基因。 结论：我们的研究结果表明，大肠杆菌和克雷伯菌属中 ESBL 和碳青霉烯酶的比例很高。在表型方法中，CDT 似乎是更好的选择，因为我国碳青霉烯酶的患病率存在很大差异。对于B类酶，CDT和mCIM/eCIM均可在常规实验室中使用。
更新日期：2020-07-01
Indian Journal of Medical Microbiology ( IF 1.4 ) Pub Date : 2020-07-01 , DOI: 10.4103/ijmm.ijmm_20_253 Varsha Gupta 1 , Meenakshi Singh 1 , Priya Datta 2 , Anku Goel 1 , Sanjay Singh 3 , Kashinath Prasad 3 , Jagdish Chander 1
Affiliation
Objective: The emergence of carbapenem-resistant Escherichia coli and Klebsiella species is a global threat. We aimed to compare two phenotypic methods and evaluate the genotypic method for the detection of beta-lactamases produced by E. coli and Klebsiella spp. Materials and Methods: One hundred and twenty-six E. coli and Klebsiella isolates were examined for phenotypic production of beta-lactamases by using disc diffusion, combined disc test (CDT) and modified carbapenem inactivation method (mCIM). All strains were also studied for the presence of various genes by polymerase chain reaction. Results: Out of 126 isolates, 96% of the isolates were extended-spectrum β-lactamase (ESBL) producers based on the presence of various ESBL genes. CDT method showed higher number of total (89%) carbapenemases in comparison to mCIM (81%). Among carbapenemases none of the isolates were Klebsiella pneumoniae carbapenemase producer by CDT, while 69% isolates were metallo-beta-lactamase (MBL) producers. Another method, mCIM/ethylene diamine tetraacetic acid mCIM showed 100% agreement for MBL detection. As regards, AmpC and class D carbapenemases; 0.04% and 16% positivity was detected, respectively, based on CDT method. Molecular analysis revealed 91% of the isolates harbouring carbapenemase genes. blaNDMwas the most common gene detected followed byblaOXA-48. Nine of the blaNDM-positive isolates also possessed blaOXA-48gene. Conclusion: Our finding shows high percentages of ESBL and carbapenemases in E. coli and Klebsiella spp. Among phenotypic methods, CDT seems to be a better choice as prevalence of carbapenemases shows lots of variation in our country. For Class B enzymes, both CDT and mCIM/eCIM can be used in the routine laboratories.
中文翻译：
大肠杆菌和克雷伯氏菌中各种 β-内酰胺酶的检测：北印度三级护理中心的一项研究
目的：耐碳青霉烯类大肠杆菌和克雷伯氏菌的出现对全球构成威胁。我们的目的是比较两种表型方法并评估用于检测大肠杆菌和克雷伯菌属产生的 β-内酰胺酶的基因型方法。材料和方法：通过使用纸片扩散、组合纸片测试 (CDT) 和改良碳青霉烯灭活方法 (mCIM)，检查 126 株大肠杆菌和克雷伯氏菌分离株的 β-内酰胺酶表型产生情况。还通过聚合酶链式反应研究了所有菌株中各种基因的存在。结果：在 126 个分离株中，基于各种 ESBL 基因的存在，96% 的分离株是超广谱 β-内酰胺酶 (ESBL) 生产者。 CDT 方法显示碳青霉烯酶总数 (89%) 高于 mCIM (81%)。根据 CDT，在碳青霉烯酶中，没有分离株是肺炎克雷伯菌碳青霉烯酶生产者，而 69% 的分离株是金属-β-内酰胺酶 (MBL) 生产者。另一种方法，mCIM/乙二胺四乙酸 mCIM 对 MBL 检测显示 100% 一致性。至于AmpC和D类碳青霉烯酶；基于CDT方法，分别检测到0.04%和16%的阳性率。分子分析显示 91% 的分离株携带碳青霉烯酶基因。 bla NDM是检测到的最常见基因，其次是bla OXA-48 。 9 个bla NDM阳性分离株也具有bla OXA-48基因。 结论：我们的研究结果表明，大肠杆菌和克雷伯菌属中 ESBL 和碳青霉烯酶的比例很高。在表型方法中，CDT 似乎是更好的选择，因为我国碳青霉烯酶的患病率存在很大差异。对于B类酶，CDT和mCIM/eCIM均可在常规实验室中使用。
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