Diagnosis and characterization of canine distemper virus through sequencing by MinION nanopore technology

Alessia Peserico,Maurilia Marcacci,Iolanda Mangone,Daniela Malatesta,Alessio Lorusso,Guendalina Zaccaria,Francesco Cirone,Marco Di Domenico,Federica Pizzurro,Annamaria Pratelli,Nicola D’Alterio,Cesare Cammà

doi:10.1038/s41598-018-37497-4

Alessia Peserico, Maurilia Marcacci + Show 10 more

Open Access

https://doi.org/10.1038/s41598-018-37497-4

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Abstract

Prompt identification of the causative pathogen of an infectious disease is essential for the choice of treatment or preventive measures. In this perspective, nucleic acids purified from the brain tissue of a dog succumbed after severe neurological signs were processed with the MinION (Oxford Nanopore Technologies, Oxford UK) sequencing technology. Canine distemper virus (CDV) sequence reads were detected. Subsequently, a specific molecular test and immunohistochemistry were used to confirm the presence of CDV RNA and antigen, respectively, in tissues. This study supports the use of the NGS in veterinary clinical practice with potential advantages in terms of rapidity and broad-range of molecular diagnosis.

Highlights

Rapid identification of the etiologic agent of an infectious disease is essential for setting up treatment and preventive measures
We described the identification and characterization of a canine distemper virus (CDV) strain from the brain tissue of a dog by using the MinION sequencing Nanopore technology
MinION real time sequencing from the brain tissue

Summary

Introduction

Rapid identification of the etiologic agent of an infectious disease is essential for setting up treatment and preventive measures. The availability of other more rapid, broad-range techniques has become more and more important in the milieu of laboratory diagnosis of infectious diseases In this context one of these novel and intriguing technologies is certainly represented by generation sequencing (NGS), even more when performed for direct identification of pathogens from clinical samples. The last large CDV outbreak in Italy was caused by a strain belonging to the Arctic lineage (prototype CDV2784/2013) It occurred in 2013 involving primarily the Abruzzi region, an area hosting several natural parks with significant animal biodiversity. In this outbreak, distemper caused overt disease in unvaccinated domestic and shepherd dogs, Apennine wolves (Canis lupus) and other wild carnivores living in natural areas of the Abruzzi and neighboring regions, including Molise. Direct diagnosis of CDV infection in animals showing neurological signs is primarily performed by molecular assays detecting the viral genome in the cerebrospinal fluid or alternatively in the urine[20,24]

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