Bialasiewicz S, Whiley D M, Buhrer-Skinner M, Bautista C, Barker K, Aitken S, Gordon R, Muller R, Lambert S B, Debattista J, Nissen M D, Sloots T P
Queensland Paediatric Infectious Diseases Laboratory, Sir Albert Sakzewski Virus Research Centre, Royal Children's Hospital and Health Service District, University of Queensland, Clinical Medical Virology Centre, Queensland, Australia.
Sex Transm Infect. 2009 Apr;85(2):102-5. doi: 10.1136/sti.2008.032607. Epub 2008 Nov 12.
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.
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.
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.
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.
本研究旨在开发一种新型尿液运输方法，用于基于自我采集的沙眼衣原体筛查。该方法需适用于沙眼衣原体的聚合酶链反应（PCR）检测，经济实惠且适合通过标准信封邮寄运输。
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颗粒状，随后在到达实验室时可重新溶解。然后使用罗氏MagNA Pure方案从重新溶解的溶液中提取DNA，用于通过PCR检测沙眼衣原体。收集了来自沙眼衣原体患病率差异很大的三个群体的尿液标本（患病率100%，n = 56；47%，n = 70；3%，n = 97）。我们使用未经处理的尿液标本和经过凝胶处理的尿液标本确定了凝胶法对沙眼衣原体PCR敏感性和特异性的影响。使用马疱疹病毒PCR检测是否存在检测抑制。
总体而言，与未经处理的尿液相比，基于凝胶的方法的敏感性范围为94.6%-100%，特异性为100%。使用经过凝胶处理的提取物未观察到PCR抑制或分析敏感性降低。
发现基于凝胶的方法适用于通过PCR检测沙眼衣原体。此外，其易于使用性、在常温下的有效性和低成本使其非常适合用于基于人群的沙眼衣原体筛查的自我采集试剂盒，特别是对于地理和社会上孤立的个体。
