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Feasibility of replacing the viral isolation technique in mice with RT-qPCR for the diagnosis of rabies
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Feasibility of replacing the viral isolation technique in mice with RT-qPCR for the diagnosis of rabies
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Journal Article
Feasibility of replacing the viral isolation technique in mice with RT-qPCR for the diagnosis of rabies
2024
Overview
Rabies is a viral encephalitis that affects mammals, including humans. Rapid and effective laboratory diagnosis of the rabies virus is critical for public health. This study evaluated the operational, technical, and financial viability of the RT-qPCR in replacement of inoculation in mice for diagnosing rabies in the laboratory routine. A total of 316 samples of mammalian brains were analyzed, 121 positives and 195 negatives, previously diagnosed by direct immunofluorescence (DIF) and mouse inoculation test (MIT). The samples were submitted to the duplex TaqMan RT-qPCR technique. The accuracy, sensitivity, and specificity of RT-qPCR were analyzed. We analyzed the costs for performing the RT-qPCR technique and compared it with the cost of MIT. The results showed 99.37% accuracy, 99.17% sensitivity, and 99.49% specificity by RT-qPCR when related to DIF and MIT results, which proved to be a robust and repeatable technique. The minimum time for a positive diagnosis was reduced in RT-qPCR (1 day) if compared to MIT (9.64 days), with a 17.7% reduction in the cost of the molecular technique. The present study demonstrated that the molecular biology technique is an efficient tool to diagnose rabies in the laboratory routine, being able to replace MIT. RESUMO: A raiva é uma encefalite viral que acomete os mamíferos, incluindo humanos. O diagnóstico laboratorial rápido e eficaz do vírus da raiva é importante para a saúde pública. Este estudo teve como objetivo avaliar a viabilidade operacional, técnica e financeira da RT-qPCR em substituição à inoculação em camundongos para o diagnóstico da raiva na rotina laboratorial. Foram analisadas 316 amostras de cérebros de mamíferos, 121 positivas e 195 negativas, previamente diagnosticadas por imunofluorescência direta (IFD) e isolamento viral em camundongos, por inoculação intracerebral em camundongos. As amostras foram submetidas à técnica duplex TaqMan RT-qPCR. A precisão, sensibilidade e especificidade da RT-qPCR foram analisadas. Os custos para realizar a técnica de RT-qPCR foram analisados e comparados com o custo do isolamento viral em camundongos. Os resultados demostraram 99,37% de acurácia, 99,17% de sensibilidade e 99,49% de especificidade pela RT-qPCR quando relacionada aos resultados de IFD e isolamento viral em camundongos, que se mostrou uma técnica robusta e com repetibilidade. O tempo mínimo para diagnóstico positivo foi reduzido na RT-qPCR (1 dia) quando comparado ao isolamento viral em camundongos (9,64 dias), e com redução de 17,7% no custo pela técnica molecular. O presente estudo demonstrou que a técnica de biologia molecular é uma ferramenta eficiente para o diagnóstico da raiva na rotina laboratorial, podendo substituir o isolamento viral em camundongos.
