Sensitive and Rapid Detection of Chlamydia trachomatis by Recombinase Polymerase Amplification Directly from Urine Samples
Sensitive and Rapid Detection of Chlamydia trachomatis by Recombinase Polymerase Amplification Directly from Urine Samples
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DOI：
10.1016/j.jmoldx.2013.08.003
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发表时间：
2014-01-01
影响因子：
4.1
通讯作者：
Langel, Olo, I
中科院分区：
文献类型：
--
作者：
Krolov, Katrin;Frolova, Jekaterina;Langel, Olo, I
沙眼衣原体是最常见的人类性传播病原体。大约三分之二的女性感染后几乎没有症状，因此常常未被诊断出来。沙眼衣原体感染是一个主要的公共卫生问题，因为它可能产生严重的长期后果，包括宫外孕、慢性盆腔疼痛和不孕的风险增加。迄今为止，已经开发了几种用于沙眼衣原体诊断的即时检测。尽管其中许多方法快速且特异性强，但它们缺乏大规模应用所需的灵敏度。我们描述了一种直接从尿液样本中进行快速、灵敏的检测。该检测使用重组酶聚合酶扩增，每次测试的最小检测限为 5 至 12 种病原体。此外，它可以在 20 分钟内直接从尿液样本中进行检测，无需在扩增反应前进行 DNA 纯化。对临床患者样本检测的初步分析特异性为 100%（95% CI，92%-100%），敏感性为 83%（95% CI，51%-97%）。整个过程相当简单，不需要特定的机械，使其可能适用于护理点环境。
Chlamydia trachomatis is the most common sexually transmitted human pathogen. Infection results in minimal to no symptoms in approximately two-thirds of women and therefore often goes undiagnosed. C. trachomatis infections are a major public health concern because of the potential severe long-term consequences, including an increased risk of ectopic pregnancy, chronic pelvic pain, and infertility. To date, several point-of-care tests have been developed for C. trachomatis diagnostics. Although many of them are fast and specific, they lack the required sensitivity for large-scale application. We describe a rapid and sensitive form of detection directly from urine samples. The assay uses recombinase polymerase amplification and has a minimum detection limit of 5 to 12 pathogens per test. Furthermore, it enables detection within 20 minutes directly from urine samples without DNA purification before the amplification reaction. Initial analysis of the assay from clinical patient samples had a specificity of 100% (95% CI, 92%-100%) and a sensitivity of 83% (95% CI, 51%-97%). The whole procedure is fairly simple and does not require specific machinery, making it potentially applicable in point-of-care settings.