一种用于定量黄热病毒的快速一步法逆转录定量聚合酶链反应

A Fast One-Step RT-qPCR to Quantify Yellow Fever Virus.

作者信息

Trindade Gisela Freitas, Fernandes Alice Gomes, de Lima Sheila Maria Barbosa

机构信息

Laboratory of Virological Technology (LATEV), Institute of Technology in Immunobiologicals (Bio-Manguinhos), Oswaldo Cruz Foundation (Fiocruz), Rio de Janeiro, Brazil.

Nuclear Magnetic Resonance (NMR) Platform, Center for Technological Development in Health (CDTS), Oswaldo Cruz Foundation (Fiocruz), Manguinhos, Rio de Janeiro, Brazil.

出版信息

Methods Mol Biol. 2025;2913:39-50. doi: 10.1007/978-1-0716-4458-4_4.

DOI:10.1007/978-1-0716-4458-4_4

PMID:40249423

Abstract

Quantitative RT-PCR (qPCR) provides a rapid, specific, and sensitive method for the determination of yellow fever viral load and presents many advantages including quantitative measurement, low contamination rate, and easy standardization. Here it is presented a semi-automatic RNA extraction followed by one-step PCR reaction. Genome copies were quantified by plasmid standard curve designed to NS5 region. This method provides a specific, rapid, and sensitive diagnosis to yellow fever virus (YFV).

摘要

定量逆转录聚合酶链反应（qPCR）为测定黄热病毒载量提供了一种快速、特异且灵敏的方法，具有许多优点，包括定量测量、低污染率和易于标准化。本文介绍了一种半自动RNA提取方法，随后进行一步PCR反应。通过针对NS5区域设计的质粒标准曲线对基因组拷贝数进行定量。该方法为黄热病毒（YFV）提供了特异、快速且灵敏的诊断。