1. Enabling animal rabies diagnostic in low-access areas: Sensitivity and specificity of a molecular diagnostic test from cerebral tissue dried on filter paper
- Author
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Rasolonjatovo, Felana, Guis, Hélène, Rajeev, Malavika, Dacheux, Laurent, Nomenjanahary, Lalaina, Razafitrimo, Girard, Rafisandrantantsoa, Jean Théophile, Cêtre-Sossah, Catherine, Heraud, Jean-Michel, Andriamandimby, Soa Fy, Institut Pasteur de Madagascar, Réseau International des Instituts Pasteur (RIIP), Animal, Santé, Territoires, Risques et Ecosystèmes (UMR ASTRE), Centre de Coopération Internationale en Recherche Agronomique pour le Développement (Cirad)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Université d'Antananarivo, Centre National de Recherche Appliquée au Développement Rural (FOFIFA), Département Systèmes Biologiques (Cirad-BIOS), Centre de Coopération Internationale en Recherche Agronomique pour le Développement (Cirad), Department of Ecology and Evolutionary Biology [Princeton], Princeton University, Centre National de Référence de la Rage - National Reference Center Rabies (CNR), Institut Pasteur [Paris], This work was funded by Institut Pasteur of Madagascar (IPM), by INTERREG FEDER TROI 2015-2017 under the DP One Health Indian Ocean (www.onehealth-oi.org) and was supported by DARRI (Institut Pasteur, Paris) and Institut Carnot 'Pasteur Microbe & Sante' (grant INNOV-36-19) and by a PTR (Programmes Tranversaux de Recherche) grant (PTR 237-19) from Institut Pasteur Paris., and Institut Pasteur [Paris] (IP)
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RNA viruses ,Viral Diseases ,[SDV]Life Sciences [q-bio] ,RC955-962 ,Artificial Gene Amplification and Extension ,L73 - Maladies des animaux ,Pathology and Laboratory Medicine ,Polymerase Chain Reaction ,Biochemistry ,Fats ,Filter Paper ,[SDV.MHEP.MI]Life Sciences [q-bio]/Human health and pathology/Infectious diseases ,Specimen Storage ,Zoonoses ,Arctic medicine. Tropical medicine ,Medicine and Health Sciences ,disease recognition [EN] ,Mammals ,Reverse Transcriptase Polymerase Chain Reaction ,Brain ,Eukaryota ,Lipids ,Laboratory Equipment ,PCR ,Infectious Diseases ,Molecular Diagnostic Techniques ,Medical Microbiology ,Viral Pathogens ,Méthode alternative ,Viruses ,Vertebrates ,Engineering and Technology ,Pathogens ,Public aspects of medicine ,RA1-1270 ,Research Article ,Neglected Tropical Diseases ,Asia ,Rabies ,Equipment ,Research and Analysis Methods ,Sensitivity and Specificity ,Microbiology ,Rage ,Specimen Handling ,Rabies Virus ,Extraction techniques ,Dogs ,Animals ,Diagnostic ,Desiccation ,Pays en développement ,Molecular Biology Techniques ,Developing Countries ,Microbial Pathogens ,Molecular Biology ,Analyse de tissus ,Organisms ,Biology and Life Sciences ,Tropical Diseases ,RNA extraction ,Storage and Handling ,Africa ,Amniotes ,[SDV.SPEE]Life Sciences [q-bio]/Santé publique et épidémiologie ,Lyssavirus - Abstract
Rabies is a lethal zoonotic encephalomyelitis that causes an estimated 59,000 human deaths yearly worldwide. Although developing countries of Asia and Africa bear the heaviest burden, surveillance and disease detection in these countries is often hampered by the absence of local laboratories able to diagnose rabies and/or the difficulties of sample shipment from low-access areas to national reference laboratories. Filter papers offer a convenient cost-effective alternative for the sampling, shipment, and storage of biological materials for the diagnosis of many pathogens including rabies virus, yet the properties of diagnostic tests using this support have not been evaluated thoroughly. Sensitivity and specificity of molecular diagnosis of rabies infection using a reverse transcription followed by a hemi-nested polymerase chain reaction (RT-hn-PCR) either directly on brain tissue or using brain tissue dried on filter paper were assessed on 113 suspected field animal samples in comparison to the direct fluorescent antibody test (FAT) recommended by the World Health Organization as one of the reference tests for rabies diagnosis. Impact of the duration of the storage was also evaluated. The sensitivity and the specificity of RT-hn-PCR i) on brain tissue were 96.6% (95% CI: [88.1–99.6]) and 92.7% (95% CI: [82.4–98.0]) respectively and ii) on brain tissue dried on filter paper 100% (95% CI: [93.8–100.0]) and 90.9% (95% CI: [80.0–97.0]) respectively. No loss of sensitivity of RT-hn-PCR on samples of brain tissue dried on filter paper left 7 days at ambient temperature was detected indicating that this method would enable analyzing impregnated filter papers sent to the national reference laboratory at ambient temperature within a 1-week shipment time. It could therefore be an effective alternative to facilitate storage and shipment of samples from low-access areas to enhance and expand rabies surveillance., Author summary Dogs are responsible for 99% of human rabies deaths. Facilitating diagnostic of rabid dogs can help further identify i) people who were exposed to rabies in order to encourage them to seek post exposure prophylaxis and ii) other exposed animals to break the transmission chain (i.e. prevent them from further transmitting the virus to other people and animals). Yet, the reference diagnostic method for animal rabies requires brain samples to be collected post-mortem and shipped under temperature-controlled conditions. The shipping of such samples is complex in low-access areas, especially in low income countries, which are often those that bear the heaviest rabies burden. Filter papers offer a convenient alternative for biological sample shipment and storage. Here the efficiency of a molecular diagnostic method applied to brain tissue dried on filter paper is compared to one of the reference methods, the direct fluorescent antibody test. Our results show it has an excellent sensitivity (it does not miss any positive samples), even when filter papers are left 7 days at ambient temperature. These results let us foresee a cost-effective alternative facilitating shipment, storage and testing of samples from rabies suspected animals from low-access areas. This could considerably enhance and expand rabies surveillance in low-income countries, allowing a more comprehensive evaluation of rabies burden, and thus reinforcing arguments for allocating funds to rabies control policies.
- Published
- 2020