Mohan Raj J R, Vittal R, Huilgol P, Bhat U, Karunasagar I
Nitte University Centre for Science Education and Research, Mangaluru, India.
National Institute of Technology Karnataka, Surathkal, Mangaluru, India.
Lett Appl Microbiol. 2018 Jul;67(1):9-14. doi: 10.1111/lam.12994. Epub 2018 May 24.
The resistance determinant bla has many variants and has been the most commonly reported gene in clinical isolates of extended spectrum beta-lactamase producing Escherichia coli. Phages have been speculated as potential reservoirs of resistance genes and efficient vehicles for horizontal gene transfer. The objective of the study was to determine the prevalence and characterize bacteriophages that harbour the resistance determinant bla . Escherichia coli specific bacteriophages were isolated from 15 samples including soil and water across Mangaluru, India using bacterial hosts that were sensitive to β-lactams. Phenotypic and genotypic characterization based on plaque morphology, host range, restriction fragment length polymorphism (RFLP), presence of bla and electron microscopy was performed. Of 36 phages isolated, seven were positive for Group 1 of bla . Based on host range and RFLP pattern, the seven phages were classified into four distinct groups, each harbouring a variant of bla . Five phages were T4-like Myoviridae by electron microscopy which was further confirmed by polymerase chain reaction (PCR) for T4 specific gp14. Generalized transduction of the CTX-M gene from these phages was also observed. The high prevalence (20%) of this gene bla in the phage pool confirms the significant role of Myoviridae members, specifically T4-like phages in the dissemination of this resistance gene.
The CTX-M gene that confers resistance to Beta-lactam class of drugs is widespread and diverse. Understanding mechanisms of antimicrobial resistance transfer is a key to devise methods for controlling it. Few studies indicate that bacteriophages are involved in the transfer of this gene but the type of phages involved and the degree of involvement remains to be explored. Our work has been able to identify the class of phages and the magnitude of involvement in the dissemination of this gene.
耐药决定簇bla有许多变体,并且一直是产超广谱β-内酰胺酶大肠杆菌临床分离株中最常报道的基因。噬菌体被推测为耐药基因的潜在储存库和水平基因转移的有效载体。本研究的目的是确定携带耐药决定簇bla的噬菌体的流行情况并对其进行特征分析。使用对β-内酰胺敏感的细菌宿主,从印度芒格洛尔的15个包括土壤和水的样本中分离出大肠杆菌特异性噬菌体。基于噬菌斑形态、宿主范围、限制性片段长度多态性(RFLP)、bla的存在情况以及电子显微镜观察进行表型和基因型特征分析。在分离出的36个噬菌体中,7个bla的第1组呈阳性。基于宿主范围和RFLP模式,这7个噬菌体被分为4个不同的组,每组都携带bla的一个变体。通过电子显微镜观察,5个噬菌体为T4样肌尾噬菌体,这通过针对T4特异性gp14的聚合酶链反应(PCR)进一步得到证实。还观察到这些噬菌体对CTX-M基因的广义转导。该bla基因在噬菌体库中的高流行率(20%)证实了肌尾噬菌体科成员,特别是T4样噬菌体在该耐药基因传播中的重要作用。
赋予对β-内酰胺类药物耐药性的CTX-M基因广泛且多样。了解抗菌药物耐药性转移机制是设计控制方法的关键。很少有研究表明噬菌体参与该基因的转移,但所涉及的噬菌体类型和参与程度仍有待探索。我们的工作能够确定参与该基因传播的噬菌体类别及其参与程度。