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使用逆转录聚合酶链反应评估临床来源分离株中参与生物膜形成的基因。

Evaluation of and Genes Involved in Biofilm Formation in Isolates from Clinical Sources Using Reverse Transcriptase PCR.

作者信息

Armoon M, Babapour E, Mirnejad R, Babapour M, Taati Moghadam M

机构信息

Biotechnology Research Center, Karaj Branch, Islamic Azad University, Karaj, Iran.

Molecular Biology Research Center, Systems biology and poisonings Institute, Baqiyatallah University of Medical Sciences, Tehran, Iran.

出版信息

Arch Razi Inst. 2024 Dec 31;79(6):1329-1335. doi: 10.32592/ARI.2024.79.6.1329. eCollection 2024 Dec.

Abstract

is recognized for its capacity to generate biofilms, which facilitate bacterial adhesion to diverse substrates and present a significant challenge to therapeutic intervention. The process of biofilm formation is dependent on the operon, with the and genes playing a pivotal role in this intricate process. The objective of this study is to investigate the role of these genes in the biofilm formation of isolates sourced from clinical settings. A total of 100 isolates were collected from clinical sources and subsequently subjected to DNA and RNA extraction using a commercial kit from Kiagen Co. To transcribe the RNA samples into cDNA, a commercial kit from Kiagen Co. was employed. The capacity to produce phenotypic and molecular biofilm formation was then measured using the microtiter plate method and PCR, respectively. The expression levels of the and genes were determined via RT-PCR (Reverse transcription polymerase chain reaction). The results indicated that 95% (95%) of the isolates were capable of producing biofilm, with 16 (16%) producing weak, 64 (64%) producing medium, and 15 (15%) producing strong biofilms. Furthermore, the gene was detected in 72% of the isolates, while the gene was detected in 58%. Of these isolates, 70 (97.2%) expressed the gene, and 53 (73.6%) expressed the gene. Conversely, four isolates (5.5%) that possessed the gene but lacked the gene did not form biofilm. One strain did not express either of the genes. The presence of either the or gene is crucial for the development of biofilm. However, further investigation is necessary to fully comprehend the intricacies of biofilm formation.

摘要

因其产生生物膜的能力而被认可,生物膜促进细菌粘附于各种底物,并对治疗干预构成重大挑战。生物膜形成过程依赖于操纵子,其中[具体基因1]和[具体基因2]基因在这一复杂过程中起关键作用。本研究的目的是调查这些基因在临床来源的[细菌名称]分离株生物膜形成中的作用。总共从临床来源收集了100株[细菌名称]分离株,随后使用凯杰公司的商业试剂盒进行DNA和RNA提取。为了将RNA样本转录成cDNA,使用了凯杰公司的商业试剂盒。然后分别使用微量滴定板法和PCR测量产生表型和分子生物膜形成的能力。通过逆转录聚合酶链反应(RT-PCR)测定[具体基因1]和[具体基因2]基因的表达水平。结果表明,95%的分离株能够产生生物膜,其中16%(16株)产生弱生物膜,64%(64株)产生中等生物膜,15%(15株)产生强生物膜。此外,72%的分离株检测到[具体基因1]基因,58%检测到[具体基因2]基因。在这些分离株中,70株(97.2%)表达[具体基因1]基因,53株(73.6%)表达[具体基因2]基因。相反,4株(5.5%)拥有[具体基因1]基因但缺乏[具体基因2]基因的分离株未形成生物膜。一株菌株两种基因均未表达。[具体基因1]或[具体基因2]基因的存在对生物膜的形成至关重要。然而,需要进一步研究以充分理解生物膜形成的复杂性。

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