利用硅基 PCR 设计用于检测白喉杆菌生物标记的白喉毒素抑制因子 (dtxR) 基因的引物

Hilari Rio Rosa Nastiti, Rachmad Bayu Kuncara
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摘要

白喉杆菌是导致白喉的细菌。白喉棒状杆菌的致病因子来自细菌产生细菌毒素的能力。毒素的产生受一组称为 tox/dtx 基因的基因调控,并受 dtxR 基因调控。本研究的目的是利用细菌 DNA 序列设计用于评估 dtxR 基因的引物。本研究是一项实验研究,采用了文献研究方法,使用了 In silico 聚合酶链式反应(PCR)、NCBI(美国国家生物技术信息中心)、Primer3Plus 和 Oligo Calculator 等应用程序。从基因库 NCBI 中获得的样本是白喉杆菌 dtxR 基因 M80337.1。使用 Primer3Plus 中新设计的引物和从 In silico PCR 数据库中提取的 50 个棒状杆菌属基因组 DNA 进行了 In silico PCR 检测。dtxR 引物对:'5-ACAGTTAGCCAAACCGTTGC-3'和 5'-TGCGTTCAACTTCGTCACTC-3' 可以产生一个 226 bp 的白喉杆菌单个 DNA 扩增片段,而其他棒状杆菌基因组没有产生扩增片段。根据研究结果,这对特异性引物可用于体外 PCR 检测,并可用于白喉的快速检测。
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Desain Primer untuk Deteksi Gen Diphtheria Toxin Repressor (dtxR) sebagai Biomarker Bakteri Corynebacterium diphtheriae Menggunakan In Silico PCR
Corynebacterium diphtheriae is the bacteria that causes diphtheria. The virulence factor of C. diphtheriae comes from the bacteria's ability to produce bacterial toxins. Toxin production is regulated by a set of genes called tox/dtx genes and is regulated by the dtxR gene. The aim of this study was to design primers used to evaluate the dtxR gene using bacterial DNA sequences. This research is experimental research with a literature study approach using the In silico Polymerase Chain Reaction (PCR), NCBI (National Center for Biotechnology Information), Primer3Plus, and Oligo Calculator applications. The sample obtained from genbank NCBI was C. diphtheriae dtxR gene M80337.1. In silico PCR examination was carried out using newly designed primers from Primer3Plus with 50 genomic DNA of Corynebacterium spp. taken from the In silico PCR database. The dtxR primer pair: '5-ACAGTTAGCCAAACCGTTGC-3' and 5'-TGCGTTCAACTTCGTCACTC-3' can produce a single DNA amplicon measuring 226 bp specifically for C. diphtheria types and no amplicon bands were generated from other Corynebacterium genomes. Based on the study results, this pair of specific primers can be used for in vitro PCR testing and can be used to develop rapid detection of diphtheria.
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