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Sensitive and Stable Molecular Detection of Dengue, Chikungunya, and Zika Viruses from Dried Blood Spots

Jaime A. Cardona-OspinaGrupo de Investigación Biomedicina, Facultad de Medicina, Institución Universitaria Visión de las Américas, Pereira, Risaralda, Colombia;
Emerging Infectious Diseases and Tropical Medicine Research Group, Sci-help, Pereira, Risaralda, Colombia;

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Victoria StittleburgDepartment of Medicine, Division of Infectious Diseases, Emory University, Atlanta, Georgia;

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Natalia Millan-BenavidezGrupo de Investigación Biomedicina, Facultad de Medicina, Institución Universitaria Visión de las Américas, Pereira, Risaralda, Colombia;

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Juliana Restrepo-ChicaGrupo de Investigación Biomedicina, Facultad de Medicina, Institución Universitaria Visión de las Américas, Pereira, Risaralda, Colombia;

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Autum KeyDepartment of Pathology and Laboratory Medicine, Emory University, Atlanta, Georgia;

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Diana Marcela Rojas-GallardoGrupo de Investigación Biomedicina, Facultad de Medicina, Institución Universitaria Visión de las Américas, Pereira, Risaralda, Colombia;

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Anne PiantadosiDepartment of Medicine, Division of Infectious Diseases, Emory University, Atlanta, Georgia;
Department of Pathology and Laboratory Medicine, Emory University, Atlanta, Georgia;

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Matthew H. CollinsDepartment of Medicine, Division of Infectious Diseases, Emory University, Atlanta, Georgia;
Department of Global Health, Rollins School of Public Health, Atlanta, Georgia

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Jesse J. WaggonerDepartment of Medicine, Division of Infectious Diseases, Emory University, Atlanta, Georgia;
Department of Global Health, Rollins School of Public Health, Atlanta, Georgia

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ABSTRACT.

Standard molecular detection of many pathogens, in particular RNA viruses, requires appropriate handling in the field for preserving the quality of the sample until processing. This could be challenging in remote tropical areas. Dengue virus (DENV), chikungunya virus (CHIKV), and Zika virus (ZIKV) are RNA viruses, prominent among the causes of fever in the tropics. We aimed to test the stability of arboviral RNA in contrived dried blood spots prepared on Whatman 903 Protein saver cards as a means of sample collection and storage. We were able to detect DENV, CHIKV, and ZIKV by real-time RT-PCR up to 180 days after card inoculation with stable Ct values across the study period. Our study supports dried blood spots (DBS) on protein saver cards as a platform for stable detection of arboviral RNA of sufficient quality to be used in diagnostic RT-PCR assays and next generation sequencing.

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Author Notes

Address correspondence to Jaime A. Cardona-Ospina, Institución Universitaria Visión de las Américas, Av. de las Américas ##98 -56, Pereira, Risaralda, Colombia, E-mail: jaime.cardona@uam.edu.co or Jesse J. Waggoner, Emory University, 1760 Haygood Dr. NE, Room E-169, Bay E-1, Atlanta, GA, E-mail: jjwaggo@emory.edu.

Financial support: This research was supported in part by the Halle Institute for Global Research and the University Research Committee of Emory University, and by the Ministerio de Ciencia, Tecnología e Innovación de Colombia, (Minciencias), Contract No 419-2020.

Authors’ addresses: Jaime A. Cardona-Ospina, Grupo de Investigación Biomedicina, Facultad de Medicina, Institución Universitaria Visión de las Américas, Pereira, Risaralda, Colombia, and Emerging Infectious Diseases and Tropical Medicine Research Group, Sci-help, Pereira, Risaralda, Colombia, E-mail: jaime.cardona@uam.edu.co. Victoria Stittleburg, Department of Medicine, Division of Infectious Diseases, Emory University, Atlanta, GA, E-mail: victoria.d.simmons@emory.edu. Natalia Millan-Benavidez, Juliana Restrepo-Chica, and Diana Marcela Rojas-Gallardo, Grupo de Investigación Biomedicina, Facultad de Medicina, Institución Universitaria Visión de las Américas, Pereira, Risaralda, Colombia, E-mails: natalia.millan@uam.edu.co, juliana.restrepoc@uam.edu.co, and diana.rojasg@uam.edu.co. Autum Key, Department of Pathology and Laboratory Medicine, Emory University, Atlanta, GA, E-mail: autum.key@emory.edu. Anne Piantadosi, Department of Medicine, Division of Infectious Diseases, Emory University, Atlanta, GA, and Department of Pathology and Laboratory Medicine, Emory University, Atlanta, GA, E-mail: anne.piantadosi@emory.edu. Matthew H. Collins and Jesse J. Waggoner, Department of Medicine, Division of Infectious Diseases, Emory University, Atlanta, GA, and Department of Global Health, Rollins School of Public Health, Atlanta, GA, Emails: matthew.collins@emory.edu and jjwaggo@emory.edu.

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