基于聚合酶链反应的肺炎克雷伯菌耐药和敏感菌株的基因分型和蛋白谱比较分析。

IF 1.9 4区生物学 Q4 BIOCHEMICAL RESEARCH METHODS

Journal of microbiological methods Pub Date : 2025-09-18 DOI:10.1016/j.mimet.2025.107265

Yadav Ranu , Srajana Nayak , Pallavi Bhat Ajakkala , Chaitra Prabhu , Caroline DSouza , Biswajit Maiti

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引用次数: 0

摘要

本研究利用基于聚合酶链反应（PCR）的基因分型方法和临床和环境来源的肺炎克雷伯菌蛋白谱，探讨了抗生素耐药和抗生素敏感分离株之间的遗传联系。研究了基于PCR的基因分型，如肠杆菌重复基因间一致性PCR （ERIC）和重复基因外回文序列PCR (Rep) PCR，以及利用外膜蛋白（OMPs）和全细胞蛋白（WCPs）进行蛋白质分析。尽管来源不同，但基于pcr的方法揭示了耐药菌株和敏感菌株之间的遗传相似性。蛋白质谱在29-43 kDa区域周围也显示出不同的条带。基于pcr的技术是可靠和可重复的。虽然OMPs和wcp谱限制了分子分型技术的选择性，但它们仍然可以帮助确定最频繁出现和最不频繁出现的蛋白带，这将有助于肺炎克伯菌分离物的分型。为了更好地了解耐药和敏感肺炎克雷伯菌分离株之间的差异，基于pcr的基因分型技术是最有效的方法。蛋白质谱和抗生素谱结果为分型过程提供了进一步的见解。基于pcr的基因分型方法是检测肺炎克雷伯菌分离株之间遗传联系的可靠资源，并与蛋白质谱分析相补充，以提高对其流行病学相关性和医院环境中抗生素耐药机制的理解。

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Comparative analysis of Klebsiella pneumoniae isolates between antibiotic-resistant and antibiotic-sensitive strains using polymerase chain reaction-based genotyping and protein profiling

This study explored the genetic connections between antibiotic-resistant and antibiotic-sensitive isolates using polymerase chain reaction (PCR) based genotyping methods and protein profiling of Klebsiella pneumoniae from clinical and environmental sources. PCR-based genotyping, such as enterobacterial repetitive intergenic consensus (ERIC) PCR and repetitive extragenic palindromic sequence-based PCR (Rep) PCR, and protein profiling using outer membrane proteins (OMPs) and whole cell proteins (WCPs) were studied. Despite their different sources, PCR-based methods revealed genetic similarity between antibiotic-resistant and sensitive isolates. The protein profile also showed varying bands around the 29–43 kDa region. The PCR-based techniques were found to be dependable and repeatable. While the OMPs and WCPs profiles limited the selectiveness of the molecular typing techniques, they could still help determine the most and least frequently occurring protein bands, which would aid in typing K. pneumoniae isolates. To better understand the differences between antibiotic-resistant and antibiotic-sensitive K pneumoniae isolates, PCR-based genotyping techniques were the most effective approach. Protein profiles and antibiogram results provided further insights into the typing process. PCR-based genotyping methods are dependable resources for examining genetic connections among K. pneumoniae isolates, supplemented by insights from protein profiling to improve understanding of their epidemiological relevance and antibiotic resistance mechanisms in hospital environments.

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来源期刊

Journal of microbiological methods 生物-生化研究方法

CiteScore

4.30

自引率

4.50%

发文量

151

审稿时长

29 days

期刊介绍： The Journal of Microbiological Methods publishes scholarly and original articles, notes and review articles. These articles must include novel and/or state-of-the-art methods, or significant improvements to existing methods. Novel and innovative applications of current methods that are validated and useful will also be published. JMM strives for scholarship, innovation and excellence. This demands scientific rigour, the best available methods and technologies, correctly replicated experiments/tests, the inclusion of proper controls, calibrations, and the correct statistical analysis. The presentation of the data must support the interpretation of the method/approach. All aspects of microbiology are covered, except virology. These include agricultural microbiology, applied and environmental microbiology, bioassays, bioinformatics, biotechnology, biochemical microbiology, clinical microbiology, diagnostics, food monitoring and quality control microbiology, microbial genetics and genomics, geomicrobiology, microbiome methods regardless of habitat, high through-put sequencing methods and analysis, microbial pathogenesis and host responses, metabolomics, metagenomics, metaproteomics, microbial ecology and diversity, microbial physiology, microbial ultra-structure, microscopic and imaging methods, molecular microbiology, mycology, novel mathematical microbiology and modelling, parasitology, plant-microbe interactions, protein markers/profiles, proteomics, pyrosequencing, public health microbiology, radioisotopes applied to microbiology, robotics applied to microbiological methods,rumen microbiology, microbiological methods for space missions and extreme environments, sampling methods and samplers, soil and sediment microbiology, transcriptomics, veterinary microbiology, sero-diagnostics and typing/identification.