Rampazzo Rita de Cássia Pontello, Zambenedetti Miriam Ribas, Alexandrino Fabiana, Jacomasso Thiago, Tschá Marcel Kruchelski, de Fillipis Ana Maria Bispo, Morello Luis Gustavo, Marchini Fabricio Klerynton
Instituto de Biologia Molecular do Paraná (IBMP), Rua Professor Algacyr Munhoz Mader, 3775 CIC, 81350-010, Curitiba, Paraná, Brazil.
Laboratório de Flavivírus, Instituto Oswaldo Cruz, Fundação Oswaldo Cruz, Avenida Brasil, 4365, Rio de Janeiro, Rio de Janeiro, Brazil.
Int J Infect Dis. 2022 Jun;119:34-37. doi: 10.1016/j.ijid.2021.12.361. Epub 2022 Jan 3.
Yellow fever (YF) is a public health threat with frequent outbreaks in tropical and subtropical areas, despite the existence of a safe and effective vaccine. The diagnosis of acute infection of the etiologic agent relies mainly on real-time reverse transcription-polymerase chain reaction (RT-qPCR)-based assays.
The aim of this study was to evaluate and compare this novel protocol for yellow fever virus (YFV) diagnosis against assays developed in-house by reference laboratories for arboviruses.
We developed a novel molecular protocol for the detection of YFV that includes an Internal Control to validate the reaction and an External Control to monitor the RNA extraction efficiency.
Our assay detects one viral genome per reaction and displays no cross-reactions with dengue (1-4), Zika, or Chikungunya viruses. This novel assay yielded 95% of agreement with the reference method recommended by the Pan American Health Organization when analyzing 204 clinical samples and cultured viruses, these samples were analyzed in 3 different diagnosis centers for arboviruses in Brazil. The data suggest the use of the proposed multiplex assay protocol to do routine tests in a clinical laboratory. This product adds higher specificity and sensitivity in addition to reduced cost per test due to hands-on time and reagent spending.
尽管存在安全有效的疫苗，但黄热病仍是一种对公共卫生构成威胁的疾病，在热带和亚热带地区频繁爆发。病原体急性感染的诊断主要依赖于基于实时逆转录聚合酶链反应（RT-qPCR）的检测方法。
本研究的目的是评估并比较这种针对黄热病毒（YFV）诊断的新方案与参考实验室针对虫媒病毒自行开发的检测方法。
我们开发了一种用于检测YFV的新型分子检测方案，其中包括用于验证反应的内控物和用于监测RNA提取效率的外控物。
我们的检测方法每个反应可检测到一个病毒基因组，并且与登革热（1-4型）、寨卡或基孔肯雅病毒无交叉反应。在分析204份临床样本和培养病毒时，这种新检测方法与泛美卫生组织推荐的参考方法的一致性为95%，这些样本在巴西的3个不同虫媒病毒诊断中心进行了分析。数据表明，建议使用这种多重检测方案在临床实验室进行常规检测。该产品除了因操作时间和试剂消耗而降低每次检测成本外，还具有更高的特异性和灵敏度。
