Tick-Borne Pathogens Screening Using a Multiplex Real-Time Polymerase Chain Reaction-Based Method.

Autor: Cardenas-Cadena SA; Molecular Medicine Laboratory, Unidad Académica de Medicina Humana y Ciencias de la Salud, Universidad Autónoma de Zacatecas, Zacatecas, 98160, México., Castañeda-Lopez ME; Molecular Medicine Laboratory, Unidad Académica de Medicina Humana y Ciencias de la Salud, Universidad Autónoma de Zacatecas, Zacatecas, 98160, México., Mollinedo-Montaño FE; Molecular Medicine Laboratory, Unidad Académica de Medicina Humana y Ciencias de la Salud, Universidad Autónoma de Zacatecas, Zacatecas, 98160, México., Vazquez-Reyes S; Molecular Medicine Laboratory, Unidad Académica de Medicina Humana y Ciencias de la Salud, Universidad Autónoma de Zacatecas, Zacatecas, 98160, México., Lara-Arias J; Orthopedics and Traumatology Service, Facultad de Medicina y Hospital Universitario 'Dr. José E. González', Universidad Autónoma de Nuevo León, Monterrey, Nuevo León, 64460, México., Marino-Martinez IA; Experimental Therapies Unit, Center for Research and Development in Health Sciences, Universidad Autónoma de Nuevo León, Monterrey, Nuevo León, 64460, México., Rodriguez-Sanchez IP; Laboratory of Molecular and Structural Physiology, Facultad de Ciencias Biológicas, Universidad Autónoma de Nuevo León, Monterrey, Nuevo León, 66455, México., Garza-Veloz I; Molecular Medicine Laboratory, Unidad Académica de Medicina Humana y Ciencias de la Salud, Universidad Autónoma de Zacatecas, Zacatecas, 98160, México., Martinez-Fierro ML; Molecular Medicine Laboratory, Unidad Académica de Medicina Humana y Ciencias de la Salud, Universidad Autónoma de Zacatecas, Zacatecas, 98160, México. margaritamf@uaz.edu.mx.
Jazyk: angličtina
Zdroj: Acta parasitologica [Acta Parasitol] 2023 Sep; Vol. 68 (3), pp. 705-710. Date of Electronic Publication: 2023 Aug 02.
DOI: 10.1007/s11686-023-00702-0
Abstrakt: Purpose: This study aims to develop and evaluate a cost-effective, user-friendly multiplex quantitative real-time polymerase chain reaction (qPCR) method for detecting multiple tick-borne pathogens associated with human and veterinary diseases.
Methods: In silico PCR was performed to design and evaluate primer sequences reported for amplifying Rickettsia spp., Borrelia spp., and Ehrlichia spp. Single and multiplex qPCR assays were then standardized to detect individual pathogens and multiple pathogens in a single reaction. Positive controls were generated to determine the dynamic range of the methods. In the validation phase, a total of 800 samples were screened for the presence of tick-borne pathogens.
Results: Identification in a single qPCR reaction (multiplex) of Ehrlichia spp., and Borrelia spp. with a limit of detection of 10 copies and Rickettsia spp. with 100 copies, a PCR efficiency (E) of 90-100% and a coefficient of correlation (R 2 ) of 0.998-0.996 for all pathogens.
Conclusion: The ability to detect three significant pathogens (Ehrlichia spp., Rickettsia spp., and Borrelia spp.) in a single qPCR reaction offers a significant advantage in the field of molecular diagnostics for tick-borne diseases. This advancement has a profound impact on public health as it facilitates the selection of appropriate treatment protocols, thereby reducing complications associated with disease progression. The streamlined approach provided by this method simplifies the diagnostic process and enables timely intervention, ultimately improving patient outcomes and mitigating the potential risks associated with untreated or misdiagnosed tick-borne infections.
(© 2023. The Author(s).)
Databáze: MEDLINE
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