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用于肠道病毒PCR检测和基因分型的粪便样本在Whatman卡中的长期保存与储存

Long-Term Preservation and Storage of Faecal Samples in Whatman Cards for PCR Detection and Genotyping of and .

作者信息

Köster Pamela Carolina, Bailo Begoña, Dashti Alejandro, Hernández-Castro Carolina, Calero-Bernal Rafael, Ponce-Gordo Francisco, González-Barrio David, Carmena David

机构信息

Parasitology Reference and Research Laboratory, Spanish National Centre for Microbiology, Majadahonda, 28220 Madrid, Spain.

Group of Parasitology, Faculty of Medicine, Corporation of Tropical Pathologies, University of Antioquia, Medellín 050010, Colombia.

出版信息

Animals (Basel). 2021 May 12;11(5):1369. doi: 10.3390/ani11051369.

Abstract

Preservation and conservation of biological specimens, including faecal samples, is a challenge in remote areas or poor-resource settings where the cold chain cannot be maintained. This study aims at evaluating the suitability of filter cards for long-term storage of faecal samples of animal and human origin positive to the diarrhoea-causing protozoan parasites, and . Three commercially available Whatman Filter Cards were comparatively assessed: the FTA Classic Card, the FTA Elute Micro Card, and the 903 Protein Saver Card. Human faecal samples positive to ( = 5) and ( = 5) were used to impregnate the selected cards at given storage (1 month, 3 months, and 6 months) periods and temperature (-20 °C, 4 °C, and room temperature) conditions. Parasite DNA was detected by PCR-based methods. Sensitivity assays and quality control procedures to assess suitability for genotyping purposes were conducted. Overall, all three Whatman cards were proven useful for the detection and molecular characterisation of and under the evaluated conditions. Whatman cards represent a simple, safe, and cost-effective option for the transportation, preservation, and storage of faecal samples without the need of the cold chain.

摘要

在偏远地区或资源匮乏地区,由于无法维持冷链,生物样本(包括粪便样本)的保存和保护是一项挑战。本研究旨在评估滤纸片对于长期储存动物和人类来源的、对引起腹泻的原生动物寄生虫呈阳性的粪便样本的适用性,以及[此处原文缺失两种寄生虫名称]。对三种市售的Whatman滤纸片进行了比较评估:FTA经典卡片、FTA洗脱微量卡片和903蛋白保存卡片。使用对[此处原文缺失两种寄生虫名称]呈阳性的人类粪便样本(每种寄生虫各5份),在给定的储存期(1个月、3个月和6个月)和温度条件(-²⁰°C、4°C和室温)下浸渍所选滤纸片。通过基于聚合酶链反应(PCR)的方法检测寄生虫DNA。进行了敏感性分析和质量控制程序,以评估用于基因分型目的的适用性。总体而言,在评估条件下,所有三种Whatman卡片都被证明可用于[此处原文缺失两种寄生虫名称]的检测和分子特征分析。Whatman卡片是一种简单、安全且经济高效的选择,可用于无需冷链的粪便样本的运输、保存和储存。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b5cc/8151430/6d6f8b307a34/animals-11-01369-g001.jpg

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