Sample adequacy controls for infectious disease diagnosis by oral swabbing.
Sample adequacy controls for infectious disease diagnosis by oral swabbing.
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DOI：
10.1371/journal.pone.0241542
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发表时间：
2020
期刊：
影响因子：
3.7
通讯作者：
Cangelosi GA
中科院分区：
文献类型：
--
作者：
Deviaene M;Weigel KM;Wood RC;Luabeya AKK;Jones-Engel L;Hatherill M;Cangelosi GA
口腔拭子正在成为诊断埃博拉、结核病、新冠肺炎等传染病的非侵入性样本类型。为了确保正确的样品采集，需要样品充足性控制（SACs）来检测口腔内采集样品的指示物质。本研究为此目的评估了两种候选sac。一种检测具有代表性的口腔微生物群（链球菌种DNA），另一种检测人类细胞（人线粒体DNA， mtDNA）。对51名健康的美国志愿者的口腔拭子（代表在口腔内收集的样本）和手拭子（代表不正确收集的样本）进行了两种目标细胞类型的定量PCR （qPCR）检测。为每个分析建立了最大化约登指数的定量周期（Cq）截止值。对于口腔拭子样本，Cq截断值≤34.9的链球菌靶点具有99.0%的敏感性和特异性，而人类mtDNA则完全区分手拭子和口拭子，Cq截断值为31.3。随后，将人类mtDNA测试应用于先前从南非结核病患者和对照组收集的口腔、舌头和牙龈拭子，以及作为阴性对照组收集的“空气拭子”（来自71名受试者的总N = 292拭子）。在这些拭子中，287/292（98%）符合预期的Cq值。在配对分析中，三个口腔部位的人类mtDNA含量难以区分，但purflock拭子比OmniSwabsTM拭子收集的人类mtDNA略多（p = 0.012）。结果表明，人类mtDNA的定量不能区分从口腔内不同部位收集的拭子。然而，它可以可靠地区分口腔拭子和非口服拭子，这使得它成为一种有用的基于口腔拭子的诊断SAC。
Oral swabs are emerging as a non-invasive sample type for diagnosing infectious diseases including Ebola, tuberculosis (TB), and COVID-19. To assure proper sample collection, sample adequacy controls (SACs) are needed that detect substances indicative of samples collected within the oral cavity. This study evaluated two candidate SACs for this purpose. One detected representative oral microbiota (Streptococcus species DNA) and the other, human cells (human mitochondrial DNA, mtDNA). Quantitative PCR (qPCR) assays for the two target cell types were applied to buccal swabs (representing samples collected within the oral cavity) and hand swabs (representing improperly collected samples) obtained from 51 healthy U.S. volunteers. Quantification cycle (Cq) cutoffs that maximized Youden’s index were established for each assay. The streptococcal target at a Cq cutoff of ≤34.9 had 99.0% sensitivity and specificity for oral swab samples, whereas human mtDNA perfectly distinguished between hand and mouth swabs with a Cq cutoff of 31.3. The human mtDNA test was then applied to buccal, tongue, and gum swabs that had previously been collected from TB patients and controls in South Africa, along with “air swabs” collected as negative controls (total N = 292 swabs from 71 subjects). Of these swabs, 287/292 (98%) exhibited the expected Cq values. In a paired analysis the three oral sites yielded indistinguishable amounts of human mtDNA, however PurFlockTM swabs collected slightly more human mtDNA than did OmniSwabsTM (p = 0.012). The results indicate that quantification of human mtDNA cannot distinguish swabs collected from different sites within the mouth. However, it can reliably distinguish oral swabs from swabs that were not used orally, which makes it a useful SAC for oral swab-based diagnosis.
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影响因子：
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作者：
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影响因子：
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