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[利用多重聚合酶链反应对唾液标本中的16S核糖体RNA和基因进行联合检测]

[Combined detection of 16S rRNA and gene in saliva specimens using multiplex PCR].

作者信息

Wang X, Sun L, Yang Z, Song S, Li N, Liu Y, Tian W, Zhao Y

机构信息

College of Medical Technology, Beihua University, Jilin 132013, China.

Reproductive Medicine Prenatal Genetics Center, Bethune First Hospital of Jilin University, Changchun 130021, China.

出版信息

Nan Fang Yi Ke Da Xue Xue Bao. 2021 Dec 20;41(12):1816-1821. doi: 10.12122/j.issn.1673-4254.2021.12.09.

Abstract

OBJECTIVE

To establish a multiplex PCR-based method for detecting (Hp) 16S rRNA gene and gene in saliva samples for investigating the prevalence of Hp in the oral cavity of Hp-infected patients with digestive tract diseases.

METHODS

Bioinformatics technique was used to design specific primers for Hp 16S rRNA and genes for Hp detection using multiplex PCR, with recombinant cloning plasmids serving as the standard positive control. Oral saliva samples were collected from 156 patients with digestive tract diseases, and Hp 16S rRNA and genes were detected using the established multiplex PCR system.

RESULTS

The established multiplex PCR system showed a strong specificity and a high sensitivity for detecting Hp 16S rRNA gene and gene, with the lowest detection limit of 103 copies/μL. The recombinant plasmids pGMT-16s and pGMT- could be used as standard positive controls for the identification of Hp. Among the 156 saliva samples, 87.2% were positive for Hp 16S rRNA gene and 23.1% for Hp gene.

CONCLUSION

Hp is highly prevalent in saliva specimens of Hp-infected patients with digestive tract diseases. The presence of Hp in the oral cavity may importantly contribute to Hp infection in the digestive tract and recurrence after treatment.

摘要

目的

建立一种基于多重聚合酶链反应(PCR)的方法,用于检测唾液样本中的幽门螺杆菌(Hp)16S rRNA基因和[具体基因名称未给出]基因,以调查消化道疾病Hp感染患者口腔中Hp的流行情况。

方法

利用生物信息学技术设计用于检测Hp 16S rRNA和[具体基因名称未给出]基因的特异性引物,采用多重PCR进行检测,以重组克隆质粒作为标准阳性对照。收集156例消化道疾病患者的口腔唾液样本,使用建立的多重PCR系统检测Hp 16S rRNA和[具体基因名称未给出]基因。

结果

建立的多重PCR系统对检测Hp 16S rRNA基因和[具体基因名称未给出]基因具有很强的特异性和高灵敏度,最低检测限为103拷贝/μL。重组质粒pGMT - 16s和pGMT - [具体基因名称未给出]可作为鉴定Hp的标准阳性对照。在156份唾液样本中,Hp 16S rRNA基因阳性率为87.2%,[具体基因名称未给出]基因阳性率为23.1%。

结论

Hp在消化道疾病Hp感染患者的唾液标本中高度流行。口腔中Hp的存在可能对消化道Hp感染及治疗后复发起重要作用。

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