Abstract

Nucleic acid-based molecular detection of mitochondrial DNA (mtDNA) has been used for the detection of eukaryotic pathogens, genetic markers, and biomedical studies in humans and animals, forensic investigation, conservation biology, and environmental monitoring. Currently available molecular mtDNA testing is predominately dependent on relatively expensive laboratory equipment, which limits test applicability for on-site field testing. Recent technical advances in molecular testing of mtDNA using microfluidic chip technology and paper-based methods, and assays based on isothermal nucleic acid amplification show promise for field assay development. Microfluidic and paper-based components for mtDNA sample preparation and isothermal amplification establish the groundwork for targeted detection. The goal is on-site field detection mtDNA assays that combine sample processing, assay setup, and visual representation of the presence of mtDNA. This review examines advantages and disadvantages of alternative field-compatible tests for on-site detection of mtDNA. Despite challenges, developing an integrated “sample-through detection” field test platform is the future direction for mtDNA molecular detection in environmental samples.

摘要

基于核酸的线粒体 DNA (mtDNA) 分子检测已被用于检测真核病原体、遗传标记和人类和动物的生物医学研究、法医调查、保护生物学和环境监测。目前可用的分子 mtDNA 测试主要依赖于相对昂贵的实验室设备，这限制了现场现场测试的测试适用性。使用微流控芯片技术和纸质方法对 mtDNA 进行分子检测的最新技术进展，以及基于等温核酸扩增的检测显示了现场检测开发的前景。用于 mtDNA 样品制备和等温扩增的微流体和纸基组件为靶向检测奠定了基础。目标是现场现场检测 mtDNA 分析，结合样本处理、分析设置和 mtDNA 存在的视觉表示。本综述探讨了用于现场检测 mtDNA 的替代现场兼容测试的优缺点。尽管存在挑战，但开发一个集成的“样品通过检测”现场测试平台是环境样品中 mtDNA 分子检测的未来方向。