Dou Maowei, Macias Natalie, Shen Feng, Bard Jennifer Dien, Domínguez Delfina C, Li Xiujun
Department of Chemistry and Biochemistry, University of Texas at El Paso, El Paso, TX, USA.
Department of Pathology and Laboratory Medicine, Children's Hospital Los Angeles, Los Angeles, California, USA.
EClinicalMedicine. 2019 Feb;8:72-77. doi: 10.1016/j.eclinm.2019.02.008. Epub 2019 Mar 5.
Pertussis is a highly contagious respiratory disease caused by the bacterium (). The infection is difficult to diagnose especially in underserved or resource-limited areas. We developed a low-cost and instrument-free diagnostic method for rapid and accurate detection of on a point-of-care (POC) testing device.
We developed a paper/polymer hybrid microfluidic biochip integrated with loop-mediated isothermal amplification (LAMP) method for the rapid and accurate detection of . This microfluidic approach was validated by testing 100 de-identified remnant clinical nasopharyngeal swabs and aspirates, which were confirmed to be either positive or negative for by a validated real-time PCR assay at the Children's Hospital Los Angeles.
The instrument-free detection results could be successfully read by the naked eye within 45 min with a limit of detection (LOD) of 5 DNA copies per well. Our optimized bacterial lysis protocol allowed the direct testing of clinical samples without any complicated sample processing/preparation (i.e. DNA extraction) or the use of any equipment (e.g. centrifuges). The validation of the microfluidic approach was accomplished by testing 100 clinical samples. High sensitivity (100%) and specificity (96%) with respect to real-time PCR were achieved.
This microfluidic biochip shows great potential for point-of-care disease diagnosis in various venues including schools and physician's offices, especially in low-resource settings in developing nations.
NIH/NIAID under award number R21AI107415, NIH RCMI Pilot Grant, the Philadelphia Foundation, the Medical Center of the Americas Foundation.
百日咳是由细菌（）引起的一种高度传染性呼吸道疾病。这种感染很难诊断，尤其是在医疗服务不足或资源有限的地区。我们开发了一种低成本且无需仪器的诊断方法，用于在即时检测（POC）设备上快速准确地检测（）。
我们开发了一种集成环介导等温扩增（LAMP）方法的纸/聚合物混合微流控生物芯片，用于快速准确地检测（）。通过检测100份匿名的剩余临床鼻咽拭子和吸出物对这种微流控方法进行了验证，这些样本在洛杉矶儿童医院通过经过验证的实时PCR检测被确认为（）阳性或阴性。
无需仪器的检测结果可在45分钟内由肉眼成功读取，检测限为每孔5个DNA拷贝。我们优化的细菌裂解方案允许直接检测临床样本，无需任何复杂的样本处理/制备（即DNA提取）或使用任何设备（如离心机）。通过检测100份临床样本完成了对微流控方法的验证。相对于实时PCR，实现了高灵敏度（100%）和高特异性（96%）。
这种微流控生物芯片在包括学校和医生办公室在内的各种场所，尤其是在发展中国家的低资源环境中，显示出用于即时疾病诊断的巨大潜力。
美国国立卫生研究院/国家过敏和传染病研究所，资助编号R21AI107415；美国国立卫生研究院RCMI试点资助；费城基金会；美洲医学中心基金会。
