A novel gel-based method for self-collection and ambient temperature postal transport of urine for PCR detection of Chlamydia trachomatis
A novel gel-based method for self-collection and ambient temperature postal transport of urine for PCR detection of Chlamydia trachomatis
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DOI：
10.1136/sti.2008.032607
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发表时间：
2009-04-01
影响因子：
3.6
通讯作者：
Sloots, T. P.
中科院分区：
文献类型：
--
作者：
Bialasiewicz, S.;Whiley, D. M.;Sloots, T. P.
目的：本研究的目的是开发一种新型尿液转运方法，用于基于自我收集的沙眼衣原体筛查。该方法需要适合沙眼衣原体PCR检测，经济且适合通过标准信封邮寄运输。方法：使用由高吸水性聚合物和缓冲剂组成的无水凝胶将尿液干燥成干燥颗粒状态，随后在到达实验室后可以重新配制。然后使用 Roche MagNA Pure 方案从重构溶液中提取 DNA，通过 PCR 检测沙眼衣原体。使用来自衣原体患病率差异很大的三个人群的尿液样本（100%，n = 56；47%，n = 70；3%，n = 97）。我们使用纯且经过凝胶处理的尿液样本确定了凝胶方法对沙眼衣原体 PCR 敏感性和特异性的影响。使用马疱疹病毒 PCR 来测试测定抑制。结果：总体而言，与纯尿液相比，基于凝胶的方法的灵敏度为 94.6-100%，特异性为 100%。使用凝胶处理的提取物未观察到 PCR 抑制或分析灵敏度降低。结论：发现基于凝胶的方法适合通过 PCR 检测沙眼衣原体。此外，其易于使用、在环境温度下有效且成本低廉，使其非常适合用于基于人群的沙眼衣原体筛查的自我收集套件，特别是对于地理和社会隔离的个体。
Objectives: The aim of this study was to develop a novel urine transport method to be used in self-collection-based screening for Chlamydia trachomatis. The method needed to be suitable for C trachomatis PCR detection, be economical and suitable for transport by standard envelope mailing.Methods: An anhydrous gel composed of super-absorbent polymer and buffering agent was used to desiccate urine into a dry granulous state, which could subsequently be reconstituted upon arrival at a laboratory. DNA was then extracted from the reconstituted solution using the Roche MagNA Pure protocol for the detection of C trachomatis by PCR. Collections of urine specimens from three populations with widely differing chlamydia prevalence (100%, n = 56; 47%, n = 70; 3%, n = 97) were used. We determined the gel method's impact on C trachomatis PCR sensitivity and specificity using neat and gel-processed urine specimens. An equine herpes virus PCR was used to test for assay inhibition.Results: Overall, the sensitivity of the gel-based method ranged from 94.6-100% compared with neat urine, with a specificity of 100%. No PCR inhibition or decrease in analytical sensitivity was observed using the gel-processed extracts.Conclusions: The gel-based method was found to be suitable for the detection of C trachomatis by PCR. In addition, its ease of use, effectiveness at ambient temperature and low cost makes it well-suited for self-collection kits used in population-based C trachomatis screening, particularly for geographically and socially isolated individuals.