Departamento de Ciencia y Tecnología, Laboratorio de Materiales Biotecnológicos (LaMaBio), Universidad Nacional de Quilmes, Bernal, Buenos Aires, Argentina.
GBEyB, Multidisciplinary Institute of Cell Biology (IMBICE, Dependent of the Argentine Research Council (CONICET), Scientific Research Commission, Province of Buenos Aires (CIC‑PBA) and UNLP), La Plata, Argentina.
Appl Microbiol Biotechnol. 2022 Sep;106(17):5729-5739. doi: 10.1007/s00253-022-12069-9. Epub 2022 Aug 2.
Micropipette tips are currently among the most used disposable devices in bioresearch and development laboratories. Their main application is the fractionation of solutions. New functionalities have recently been added to this device, widening their applications. This paper analyzed disposable micropipette tips as reagent holders of PCR reagents. PCR has become a prevalent and often indispensable technique in biological laboratories for various applications, such as the detection of coronavirus and other infectious diseases. A functional micropipette tip was implemented to simplify PCR analysis and reduce the contamination chances of deoxynucleotides and specific primers. This disposable device is prepared by tip coating processes of reagents, using polyvinyl alcohol polymer and additives. The coated layer is optimized to load and release PCR reagents efficiently. As a proof of concept, we show that the detection of Bordetella pertussis, the etiological agent of whooping cough whose diagnostic relies on PCR, can be quickly done using practical-functional tips. This device is an excellent example of testing the functionality and contribution of molecular diagnostic PCR tips. KEY POINTS: • Functional micropipette tips are prepared by coating with dNTPs and primers. • Functional tips are used to replace dNTPs and primers in the PCR master mix. • PCR diagnostic of Bordetella pertussis is performed using functional tips.
目前，微量移液器吸头是生物研究和开发实验室中使用最广泛的一次性设备之一。其主要应用是溶液的分馏。最近，该设备增加了新的功能，扩大了其应用范围。本文将一次性微量移液器吸头分析为 PCR 试剂的试剂架。PCR 已成为生物实验室中用于各种应用的流行且通常必不可少的技术，例如检测冠状病毒和其他传染病。实施了一种功能齐全的微量移液器吸头，以简化 PCR 分析并减少脱氧核苷酸和特定引物的污染机会。该一次性设备是通过试剂的尖端涂层工艺制备的，使用聚乙烯醇聚合物和添加剂。优化了涂层以有效加载和释放 PCR 试剂。作为概念验证，我们展示了可以使用实用功能型吸头快速进行百日咳的病原体博德特氏菌（百日咳的病原体，其诊断依赖于 PCR）的检测。该设备是测试分子诊断 PCR 吸头功能和贡献的一个极好范例。要点： • 通过用 dNTP 和引物涂覆来制备功能性微量移液器吸头。 • 功能性吸头用于替代 PCR 主混合物中的 dNTP 和引物。 • 使用功能性吸头进行百日咳博德特氏菌的 PCR 诊断。
