Validation of a diagnostic tool for the diagnosis of lumpy skin disease

Background Lumpy skin disease (LSD) is caused by LSD virus which is a member of the Capripoxvirus (CaPV) genus. Although PCR provides for a rapid and sensitive diagnosis, it has limited use due to its complexity in terms of cost, time and equipment. Loop‐mediated isothermal amplification (LAMP) is a...

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Veröffentlicht in:Veterinary dermatology 2018-12, Vol.29 (6), p.532-e178
Hauptverfasser: Mwanandota, Julius J., Macharia, Mercy, Ngeleja, Chanasa M., Sallu, Raphael S., Yongolo, Mmeta G., Mayenga, Charles, Holton, Timothy A.
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Sprache:eng
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Zusammenfassung:Background Lumpy skin disease (LSD) is caused by LSD virus which is a member of the Capripoxvirus (CaPV) genus. Although PCR provides for a rapid and sensitive diagnosis, it has limited use due to its complexity in terms of cost, time and equipment. Loop‐mediated isothermal amplification (LAMP) is a simple, specific and cost‐effective method with a diagnostic accuracy similar to PCR. Objectives/Hypothesis To compare the detection rate (DR) of two LAMP assays versus PCR for the detection of CaPV. Animals This study used 105 apparently health animals (AHA) and 59 clinically sick animals (CSA). Methods and materials PCR and LAMP assays (LAMP1 and LAMP 2) were compared for detection of CaPV from AHA and CSA using blood and tissue samples. The detection was confirmed by sequencing of PCR positive samples. Analytical sensitivity and specificity of LAMP assays also were assessed. Results The DR in CSA was 13.6% for PCR whereas for LAMP it was 39.0% and 25.4% for LAMP 1 and 2 methods, respectively. In AHA, the LAMP assay DR was 14.3% and 1.9% for LAMP 1 and 2, respectively. Phylogenetic tree analysis confirmed the identity of CaPV. Analytic sensitivity showed a detection limit of 8 copies/μL. The analytic specificity test showed no cross detection with other infectious agents. Conclusion and clinical importance Good sensitivity and specificity results for LAMP assay support its application in the routine diagnosis of LSD, whereas its ability to detect LSDV in apparently healthy animals shows its usefulness in identifying populations at risk of LSD. Résumé Contexte La dermatose nodulaire contagieuse (LSD) est due à un virus LSD du genre Capripoxvirus (CaPV). Bien que la PCR fournisse un diagnostic sensible et rapide, son utilisation est limitée en raison de sa complexité, de son coût, du temps et de l’équipement. La LAMP (Loop‐Mediated Isothermal Amplification) est une méthode simple, spécifique et peu couteuse avec un diagnostic aussi précis qu'une PCR. Objectifs/Hypothèses Comparer le taux de détection (DR) de deux tests LAMP contre PCR pour la détection de CaPV. Sujets Cette étude utilise 105 animaux cliniquement sains (AHA) et 59 animaux cliniquement atteints (CSA). Matériel et méthode Les tests PCR et LAMP (LAMP1 et LAMP2) ont été comparés pour la détection de CaPV de AHA et CSA à l'aide d’échantillons de tissu et de sang. La détection a été confirmée par séquençage d’échantillon à PCR positive. La sensibilité et la spécificité des tests LAMP ont également été
ISSN:0959-4493
1365-3164
DOI:10.1111/vde.12690