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一项比较传统PCR和实时PCR检测尿液及粪便样本中粪类圆线虫的诊断研究。

A diagnostic study comparing conventional and real-time PCR for Strongyloides stercoralis on urine and on faecal samples.

作者信息

Formenti Fabio, La Marca Giulia, Perandin Francesca, Pajola Barbara, Romano Miryam, Santucci Beatrice, Silva Ronaldo, Giorli Giovanni, Bisoffi Zeno, Buonfrate Dora

机构信息

Centre for Tropical Diseases, IRCCS Sacro Cuore Don Calabria Hospital, Negrar, 37024, Italy.

Centre for Tropical Diseases, IRCCS Sacro Cuore Don Calabria Hospital, Negrar, 37024, Italy.

出版信息

Acta Trop. 2019 Feb;190:284-287. doi: 10.1016/j.actatropica.2018.12.001. Epub 2018 Dec 3.

Abstract

Strongyloides stercoralis is a soil-transmitted helminth with a wide distribution in tropical and subtropical areas. The diagnosis of S. stercoralisinfection can be challenging, due to the low sensitivity of microscopic examination of stool samples and coproculture. In the last decade, different in-house molecular biology techniques for S. stercoralis have been implemented. They demonstrated good accuracy, although sensitivity does not seem sufficiently high yet. Recently, a novel PCR technique has been evaluated for the detection of S. stercoralis DNA in urine. Aim of this work was to compare the sensitivity of the real-time PCR (qPCR) on feces routinely used at the Centre for Tropical Disease (CTD) of Negrar, Verona, Italy, with that of the novel based PCR on urine. As secondary objective, we evaluated a Urine Conditioning Buffer ® (Zymoresearch) with the aim of improving nucleic acid stability in urine during sample storage/transport at ambient temperatures. Patients attending the CTD and resulting positive at routine screening with serology for S. stercoralis were invited, previous written consent, to supply stool and urine samples for molecular biology. A convenience sample of 30 patients was included. The sensitivity of qPCR on feces resulted 63%, and that of based PCR on urine was 17%. In all the samples treated with the Urine Conditioning Buffer ® there was no detectable DNA. In conclusion, the sensitivity of the novel technique resulted low, and needs further implementation before being considered as a valid alternative to the validated method.

摘要

粪类圆线虫是一种土源性蠕虫,在热带和亚热带地区广泛分布。由于粪便样本显微镜检查和粪便培养的敏感性较低,粪类圆线虫感染的诊断可能具有挑战性。在过去十年中,已实施了不同的针对粪类圆线虫的内部分子生物学技术。尽管敏感性似乎还不够高,但这些技术显示出良好的准确性。最近,一种新型PCR技术已被评估用于检测尿液中的粪类圆线虫DNA。这项工作的目的是比较意大利维罗纳内格拉尔热带病中心(CTD)常规使用的粪便实时PCR(qPCR)与基于尿液的新型PCR的敏感性。作为次要目标,我们评估了一种尿液处理缓冲液®(Zymoresearch),目的是在常温下样本储存/运输期间提高尿液中核酸的稳定性。邀请在CTD就诊且血清学常规筛查粪类圆线虫呈阳性的患者,在获得书面同意后,提供粪便和尿液样本用于分子生物学检测。纳入了30名患者的便利样本。粪便qPCR的敏感性为63%,基于尿液的PCR的敏感性为17%。在所有用尿液处理缓冲液®处理的样本中均未检测到DNA。总之,新技术的敏感性较低,在被视为经过验证方法的有效替代方法之前,还需要进一步改进。

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