利用管家基因检测人类粪便样品中的弧菌

Aysan Karamghoshchi, A. Ahmadi, M. Arjomandzadegan, M. Akbari, Elahe Ghorbani Marghmaleki
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摘要

背景:Arcobacter是人类和牲畜中最常见的细菌之一,可导致人类肠胃炎以及动物生殖器和肠道疾病。这种细菌被认为是腹泻的主要原因。在分子研究中,16SrRNA基因主要作为测定Arcobacter的标准基因。本研究的目的是比较glyA、atpA和gyrA基因与16srrna基因对Arcobacter的分子检测。方法:采用标本库中从患者和健康人的粪便标本中分离的61份Arcobacter DNA。为了检测Arcobacter,使用16SrRNA以及glyA、atpA和gyrA基因的引物进行聚合酶链反应(PCR)。对PCR得到的产物进行测序。结果:增殖反应的结果表明所设计的引物和相关分子实验的准确性。结果显示,61份glyA基因样品中Arcobacter阳性检出率为65.57%。这一比例高于16SrRNA(42.62%)、gyrA(42.62%)和atpA(24.59%)。分析结果具有统计学意义。结论:考虑到大多数细菌的16SrRNA中存在重复序列,研究人员对结果的解释可能很困难。本研究结果表明,glyA基因对Arcobacter的诊断比其他基因更敏感和准确。在Arcobacter的诊断研究中,glyA基因被认为是16SrRNA的替代品。
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Molecular Detection of Arcobacter in Human Stool Samples Using Housekeeping Genes
Background: Arcobacter is one of the most common bacteria in humans and livestock, leading to gastroenteritis in humans as well as genital and enteric diseases in animals. This bacterium is known to be the main cause of diarrhea. In molecular studies, the 16SrRNA gene was primarily used as the standard gene for the determination of the Arcobacter. The purpose of this study was to investigate the molecular detection of Arcobacter using glyA, atpA, and gyrA genes compared to16SrRNA. Methods: In this study, 61 samples of Arcobacter DNA isolated from fecal specimens of patients and healthy individuals in the sample bank were used. In order to detect Arcobacter, the intended primers for 16SrRNA as well as glyA, atpA, and gyrA genes were used for polymerase chain reaction (PCR). The products obtained from the PCR were sequenced. Results: The results of the proliferation reactions indicated the accuracy of the intended primers and the associated molecular experiments. Our results showed that 65.57% of the cases were detected to be positive for Arcobacter among 61 samples using the glyA gene. This percentage was higher compared to 16SrRNA (42.62%), gyrA (42.62%), and atpA (24.59%). The analysis was statistically significant. Conclusions: Given the presence of repetitive sequences in the 16SrRNA in most bacteria, the interpretation of the results is likely to be difficult for researchers. The results of this study showed more sensitivity and accurate diagnosis of Arcobacter using the glyA gene than other studied genes. In diagnostic studies of Arcobacter, the glyA gene is proposed as an alternative to the 16SrRNA.
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