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[应用巢式聚合酶链反应检测人唾液中的幽门螺杆菌]

[Detection of Helicobacter pylori in human saliva by using nested polymerase chain reaction].

作者信息

Song M

机构信息

Dept. of Histology and Embryology, First Medical University of PLA, Guangzhou.

出版信息

Zhonghua Liu Xing Bing Xue Za Zhi. 1993 Aug;14(4):237-40.

PMID:8143325
Abstract

Epidemiologic studies suggest person to person spread of Helicobacter pylori (H. pylori), but the exact mechanism is unknown. Spread through oral secretions has been suggested, however, it has proved very difficult to grow the organism from areas outside the stomach. A nested polymerase chain reaction (N-PCR) for the specific detection of H. pylori was developed with two primer pairs (nested primers) derived from the urease gene A of H. pylori. The N-PCR could detect all 21 H. pylori strains, including 20 isolated strains and 1 reference strain NCTC 14126, but could not detect other bacterial species, showing the N-PCR assay to be 100% specific. Tenfold serial dilution experiments revealed the detection of as little as 0.1 fg DNA by N-PCR. To evaluate the PCR assay for clinical samples, gastric biopsy and aspirate specimens were tested by N-PCR, and the results were compared with those of culture, urease test and histologic examination (reference standard, RS). Among 57 biopsy specimens, H. pylori sequence was detected by PCR in 39 of 39 (100%) positive tissues and in none of 18 negative tissues. H-pylori was detected in saliva of 11 out of 19 cases in which H. pylori was positive in gastric mucosa by PCR. Whereas, PCR was positive in saliva of only one out of 8 cases in which H. pylori was negative in gastric mucosa. Six gastric aspirate specimens were positive by N-PCR. PCR is an accurate and sensitive method that can detect the presence of H. pylori without the need of culture.

摘要

流行病学研究表明幽门螺杆菌(H. pylori)可在人与人之间传播,但确切机制尚不清楚。曾有人提出通过口腔分泌物传播,然而,要从胃外区域培养出这种微生物却非常困难。利用源自幽门螺杆菌脲酶基因A的两对引物(巢式引物)开发了一种用于特异性检测幽门螺杆菌的巢式聚合酶链反应(N-PCR)。N-PCR能够检测所有21株幽门螺杆菌菌株,包括20株分离菌株和1株参考菌株NCTC 14126,但无法检测其他细菌种类,表明N-PCR检测具有100%的特异性。十倍系列稀释实验显示N-PCR能检测低至0.1 fg的DNA。为评估该PCR检测方法对临床样本的效果,对胃活检和抽吸标本进行了N-PCR检测,并将结果与培养、尿素酶试验及组织学检查(参考标准,RS)的结果进行比较。在57份活检标本中,PCR在39份(100%)阳性组织中检测到幽门螺杆菌序列,在18份阴性组织中均未检测到。在19例胃黏膜中幽门螺杆菌PCR检测呈阳性的病例中,有11例在唾液中检测到幽门螺杆菌。而在8例胃黏膜中幽门螺杆菌阴性的病例中,只有1例唾液PCR检测呈阳性。6份胃抽吸标本N-PCR检测呈阳性。PCR是一种准确且灵敏的方法,无需培养就能检测幽门螺杆菌的存在。

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引用本文的文献

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Molecular biology of microbial ureases.
Microbiol Rev. 1995 Sep;59(3):451-80. doi: 10.1128/mr.59.3.451-480.1995.

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