Malik Muqtadir, Kumar Mahadevan, Bhalla Gurpreet Singh, Tandel Kundan
Graded Specialist (Microbiology), INHS Asvini, Colaba, Mumbai, India.
Professor (Microbiology), Bharati Vidyapeeth University Medical College, Pune, India.
Med J Armed Forces India. 2024 Dec;80(Suppl 1):S223-S231. doi: 10.1016/j.mjafi.2023.09.007. Epub 2023 Nov 11.
This study aims to enhance the existing knowledge of the prevalence of genes responsible for beta-lactam resistance and aminoglycoside resistance in gram negative organisms by molecular detection of extended spectrum beta-lactamase and aminoglycoside modifying enzymes in multidrug-resistant gram-negative bacteria.
Out of 864 gram-negative isolates, 710 were phenotypically identified as multidrug-resistant by antibiotic susceptibility testing. From the above isolates, 102 representative isolates as per sample size calculated were selected for further molecular studies. The presence of blaTEM, blaCTX-M blaSHV, and five AmpC genes was detected by real-time polymerase chain reaction (PCR). Conventional PCR was performed to detect seven aminoglycoside modifying enzyme genes namely aac(6')-Ib, aac(6')-Ic, aac(3)-Ia, aac(3)-Ib, aac(3)-IIa, ant(2'')-Ia, and ant(4'')-IIa.
Most common multidrug-resistant isolate was Klebsiella pneumoniae (35%) followed by Escherichia coli (30%). Among the 102 selected isolates all harboured blaTEM gene, 71 (69.6%) harboured blaCTX-M gene and 48 (47%) blaSHV gene. Among the selected isolates 60% showed the presence of AmpC genes. Most common aminoglycosie modifying enzyme gene was AAC 6' Ib (51%) followed by ANT 2" Ia (36%).
This study suggests a wider use of molecular methods using specific PCR amplification of resistance genes. It would be beneficial to perform the molecular identification of antimicrobial resistance genes to effectively monitor and manage antibiotic resistance, administer appropriate antimicrobial medication, practice antimicrobial stewardship and improve hospital infection control procedures.
本研究旨在通过对多重耐药革兰氏阴性菌中的超广谱β-内酰胺酶和氨基糖苷类修饰酶进行分子检测,增进对革兰氏阴性菌中负责β-内酰胺耐药性和氨基糖苷类耐药性基因流行情况的现有认识。
在864株革兰氏阴性分离株中,通过抗生素敏感性试验,有710株在表型上被鉴定为多重耐药。从上述分离株中,根据计算出的样本量选择102株代表性分离株进行进一步的分子研究。通过实时聚合酶链反应(PCR)检测blaTEM、blaCTX-M、blaSHV和五种AmpC基因的存在。进行常规PCR以检测七种氨基糖苷类修饰酶基因,即aac(6')-Ib、aac(6')-Ic、aac(3)-Ia、aac(3)-Ib、aac(3)-IIa、ant(2'')-Ia和ant(4'')-IIa。
最常见的多重耐药分离株是肺炎克雷伯菌(35%),其次是大肠杆菌(30%)。在102株选定的分离株中,所有分离株都携带blaTEM基因,71株(69.6%)携带blaCTX-M基因,48株(47%)携带blaSHV基因。在选定的分离株中,60%显示存在AmpC基因。最常见的氨基糖苷类修饰酶基因是AAC 6' Ib(51%),其次是ANT 2" Ia(36%)。
本研究表明,使用针对耐药基因进行特异性PCR扩增的分子方法具有更广泛的用途。对抗菌药物耐药基因进行分子鉴定将有助于有效监测和管理抗生素耐药性、合理使用抗菌药物、实施抗菌药物管理并改进医院感染控制程序。