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使用实时聚合酶链式反应对溃疡性结肠炎患者和非溃疡性结肠炎患者的三种肠道微生物群进行定量分析。

Quantitative analysis of the three gut microbiota in UC and non-UC patients using real-time PCR.

作者信息

Al-Bayati Luma, Nayeri Fasaei Bahar, Merat Shahin, Bahonar Alireza, Ghoddusi Arefeh

机构信息

Department of Microbiology and Immunology, Faculty of Veterinary Medicine, University of Tehran, Tehran, Iran; Department of Microbiology, Faculty of Medicine, University of Wassit, Iraq.

Department of Microbiology and Immunology, Faculty of Veterinary Medicine, University of Tehran, Tehran, Iran.

出版信息

Microb Pathog. 2023 Aug;181:106198. doi: 10.1016/j.micpath.2023.106198. Epub 2023 Jun 8.

Abstract

BACKGROUND

and study aims: Gastrointestinal microbiota are closely related to the pathogenesis of ulcerative colitis (UC). This study aimed at quantification of F. prausnitzii, Provetella, and Peptostreptococcus in UC and non-UC patients using Real-Time PCR and a new set of primers were also validated for this purpose.

MATERIALS AND METHODS

In this study, the relative abundance of microbial populations between the UC and non-UC subjects were evaluated by quantitative real-time polymerase chain reaction (qRT-PCR). DNA extraction from biopsies and polymerase chain reaction (PCR) amplification of bacterial 16S rRNA gene-targeted species-specific primers was performed to detect the anaerobic bacterial species. The qRT-PCR was used to show the relative change in the bacterial populations of F. prausnitzii, Provetella, and Peptostreptococcus in the UC and non-UC subjects.

RESULTS

Our data for detection of the anaerobic intestinal flora showed Faecalibacterium prausnitzii, Provetella and Peptostreptococcus were the predominant microflora in the controls and showed significant differences (p = 0.002, 0.025 and 0.039, respectively). The qRT-PCR analyses of F. prausnitzii, Provetella and Peptostreptococcus were 8.69-, 9.38- and 5.77-higher, respectively, in the control group than in the UC group.

CONCLUSION

The results of this study showed decreased abundance of F. prausnitzii, Provetella and Peptostreptococcus in the intestine of UC patients in comparison to non-UC patients. Quantitative RT-PCR, as a progressive and sensitive method, could be useful for evaluation of bacterial populations in patients with inflammatory bowel diseases to attain appropriate therapeutic strategies.

摘要

背景与研究目的

胃肠道微生物群与溃疡性结肠炎(UC)的发病机制密切相关。本研究旨在通过实时聚合酶链反应(Real-Time PCR)对UC患者和非UC患者中的普拉梭菌、普氏菌属和消化链球菌进行定量分析,并为此验证了一组新引物。

材料与方法

在本研究中,通过定量实时聚合酶链反应(qRT-PCR)评估UC患者和非UC患者之间微生物群的相对丰度。从活检组织中提取DNA,并使用针对细菌16S rRNA基因的物种特异性引物进行聚合酶链反应(PCR)扩增,以检测厌氧细菌种类。qRT-PCR用于显示UC患者和非UC患者中普拉梭菌、普氏菌属和消化链球菌细菌群的相对变化。

结果

我们检测厌氧肠道菌群的数据显示,普拉梭菌、普氏菌属和消化链球菌是对照组中的主要微生物群,且差异显著(分别为p = 0.002、0.025和0.039)。对照组中普拉梭菌、普氏菌属和消化链球菌的qRT-PCR分析结果分别比UC组高8.69倍、9.38倍和5.77倍。

结论

本研究结果表明,与非UC患者相比,UC患者肠道中普拉梭菌、普氏菌属和消化链球菌的丰度降低。定量RT-PCR作为一种先进且灵敏的方法,可用于评估炎症性肠病患者的细菌群,以制定合适的治疗策略。

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