Use of saliva-based qPCR diagnostics for the accurate, rapid, and inexpensive detection of strep throat.

IF 2.2 Q2 MEDICINE, GENERAL & INTERNAL

Diagnosis Pub Date : 2024-01-05 eCollection Date: 2024-05-01 DOI:10.1515/dx-2023-0134

Madeline H Peachey, Kristopher E Kubow, Kristina B Blyer, Julia A Halterman

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引用次数: 0

Abstract

Objectives: Outpatient health care facilities are essential for quickly diagnosing common infectious diseases such as bacterial and viral pharyngitis. The only form of pharyngitis requiring antibiotics is strep throat (ST); however, antibiotic prescription rates are much higher than ST prevalence, suggesting antibiotics are being inappropriately prescribed. Current rapid ST diagnostics may be contributing to this problem due to the low sensitivity and variable specificity of these tests. It is best practice to verify a negative ST diagnosis with a group A Streptococcus (GAS) culture, but many clinics do not perform this test due to the additional cost and 24-72 h required to obtain results. This indicates there is great need for more accurate rapid diagnostic tools in outpatient facilities. We hypothesized that next generation qPCR technology could be adapted to detect GAS DNA from saliva samples (instead of the traditional throat swab) by creating a simple, fast, and inexpensive protocol.

Methods: Saliva specimens collected from patients at James Madison University Health Center were used to test the effectiveness of our Chelex 100-based rapid DNA extraction method, followed by a fast protocol developed for the Open qPCR machine to accurately detect ST.

Results: Our final saliva processing and qPCR protocol required no specialized training to perform and was able to detect ST with 100 % sensitivity and 100 % specificity (n=102) in 22-26 min, costing only $1.12 per sample.

Conclusions: Saliva can be rapidly analyzed via qPCR for the accurate and inexpensive detection of ST.

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利用基于唾液的 qPCR 诊断技术，准确、快速、低成本地检测链球菌咽喉炎。

目的：门诊医疗机构对于快速诊断细菌性和病毒性咽炎等常见传染病至关重要。唯一需要使用抗生素的咽炎是链球菌性咽喉炎（ST）；然而，抗生素处方率远远高于 ST 发病率，这表明抗生素处方不当。目前的快速 ST 诊断方法可能是造成这一问题的原因之一，因为这些检测方法的灵敏度低且特异性不一。最佳做法是用 A 组链球菌（GAS）培养来验证 ST 阴性诊断结果，但由于费用昂贵且需要 24-72 小时才能获得结果，许多诊所并不进行这种检测。这表明门诊机构亟需更准确的快速诊断工具。我们推测，下一代 qPCR 技术可用于检测唾液样本（而非传统的咽拭子）中的 GAS DNA，方法简单、快速且成本低廉：方法：从詹姆斯-麦迪逊大学健康中心的患者唾液样本中收集的样本被用来测试我们基于 Chelex 100 的快速 DNA 提取方法的有效性，然后使用为 Open qPCR 仪器开发的快速方案来准确检测 ST：结果：我们最终的唾液处理和 qPCR 方案无需专业培训即可执行，并能在 22-26 分钟内检测出 ST，灵敏度和特异性均为 100%（n=102），每个样本的成本仅为 1.12 美元：结论：通过 qPCR 可以快速分析唾液，准确、低成本地检测 ST。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Diagnosis MEDICINE, GENERAL & INTERNAL-

CiteScore

7.20

自引率

5.70%

发文量

期刊介绍： Diagnosis focuses on how diagnosis can be advanced, how it is taught, and how and why it can fail, leading to diagnostic errors. The journal welcomes both fundamental and applied works, improvement initiatives, opinions, and debates to encourage new thinking on improving this critical aspect of healthcare quality.　 Topics: -Factors that promote diagnostic quality and safety -Clinical reasoning -Diagnostic errors in medicine -The factors that contribute to diagnostic error: human factors, cognitive issues, and system-related breakdowns -Improving the value of diagnosis – eliminating waste and unnecessary testing -How culture and removing blame promote awareness of diagnostic errors -Training and education related to clinical reasoning and diagnostic skills -Advances in laboratory testing and imaging that improve diagnostic capability -Local, national and international initiatives to reduce diagnostic error