Detection of Streptococcus anginosus in fecal samples using PCR-CRISPR /Cas12a system.

IF 1.8 4区医学 Q3 BIOCHEMICAL RESEARCH METHODS

Bioanalysis Pub Date : 2025-06-01 Epub Date: 2025-06-12 DOI:10.1080/17576180.2025.2518042

Yan Zheng, BiXia Liu, QianFei Zuo, Fei Deng, ChunHui Lan

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引用次数: 0

Abstract

Objective: To develop a highly sensitive and specific detection method based on PCR-CRISPR/Cas12a for the detection of Streptococcus anginosus (S. anginosus) in feces and to evaluate its detection rate in the general population as well as its potential as a gastrointestinal tumor marker.

Materials and methods: Specific primers and crRNA targeting the 16S rDNA of S. anginosus were designed to construct a PCR-CRISPR/Cas12a detection system. A total of 230 fecal samples were collected from the general population, and bacterial DNA was extracted. The target gene was detected using this system to verify its sensitivity, specificity, and stability.

Results: The established detection system demonstrated strong specificity, with stable recognition of S. anginosus, and a minimum detection limit of 10^-7 ng/μL. The detection rate of S. anginosus in fecal samples from the general population was 51.7%.

Conclusion: The PCR-CRISPR/Cas12a system can efficiently detect S. anginosus in feces, providing a reliable technical tool for exploring its association with gastrointestinal tumors.

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利用PCR-CRISPR /Cas12a系统检测粪便标本中的血管链球菌。

目的：建立一种基于PCR-CRISPR/Cas12a技术检测粪便中血管链球菌（S. anginosus）的高灵敏度和特异性检测方法，评估其在普通人群中的检出率及作为胃肠道肿瘤标志物的潜力。材料与方法：设计特异性引物和靶向血管棘鱼16S rDNA的crRNA，构建PCR-CRISPR/Cas12a检测体系。从普通人群中收集了230份粪便样本，提取了细菌DNA。利用该系统检测目的基因，验证其敏感性、特异性和稳定性。结果：所建立的检测体系特异性强，对血管葡萄球菌识别稳定，最低检出限为10 ~ 7 ng/μL。一般人群粪便标本中血管链球菌检出率为51.7%。结论：PCR-CRISPR/Cas12a系统可有效检测粪便中的血管链球菌，为探索其与胃肠道肿瘤的相关性提供了可靠的技术工具。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Bioanalysis BIOCHEMICAL RESEARCH METHODS-CHEMISTRY, ANALYTICAL

CiteScore

3.30

自引率

16.70%

发文量

审稿时长

2 months

期刊介绍： Reliable data obtained from selective, sensitive and reproducible analysis of xenobiotics and biotics in biological samples is a fundamental and crucial part of every successful drug development program. The same principles can also apply to many other areas of research such as forensic science, toxicology and sports doping testing. The bioanalytical field incorporates sophisticated techniques linking sample preparation and advanced separations with MS and NMR detection systems, automation and robotics. Standards set by regulatory bodies regarding method development and validation increasingly define the boundaries between speed and quality. Bioanalysis is a progressive discipline for which the future holds many exciting opportunities to further reduce sample volumes, analysis cost and environmental impact, as well as to improve sensitivity, specificity, accuracy, efficiency, assay throughput, data quality, data handling and processing. The journal Bioanalysis focuses on the techniques and methods used for the detection or quantitative study of analytes in human or animal biological samples. Bioanalysis encourages the submission of articles describing forward-looking applications, including biosensors, microfluidics, miniaturized analytical devices, and new hyphenated and multi-dimensional techniques. Bioanalysis delivers essential information in concise, at-a-glance article formats. Key advances in the field are reported and analyzed by international experts, providing an authoritative but accessible forum for the modern bioanalyst.