Detection of canine distemper virus in dogs by real-time RT-PCR

Elia, Gabriella; Decaro, Nicola; Martella, Vito; Cirone, Francesco; Lucente, Maria Stella; Lorusso, Eleonora; Trani, Livia Di; Buonavoglia, Canio

doi:10.1016/j.jviromet.2006.05.004

Cited by 191 publications

(201 citation statements)

References 14 publications

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“…This poses a problem from a diagnostic stand point, given that CDV MLV vaccine can be detected in the blood, urine, and swabs by highly sensitive real-time reverse transcription polymerase chain reaction (RT-PCR) tests, including the one routinely used in the University of Tennessee, College of Veterinary Medicine, Clinical Virology Diagnostic Laboratory (Knoxville, Tennessee). 7 This vaccine interference is recognized to limit the effectiveness of a real-time RT-PCR test for diagnosis of CDV and has prompted a commercial laboratory to develop a test a to deal with this problem. This test provides a quantitative measure of the CDV viral load.…”

Section: Introductionmentioning

confidence: 99%

Real-time reverse transcription polymerase chain reaction method for detection of Canine distemper virus modified live vaccine shedding for differentiation from infection with wild-type strains

et al. 2014

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Abstract. Canine distemper virus (CDV) remains a common cause of infectious disease in dogs, particularly in highdensity housing situations such as shelters. Vaccination of all dogs against CDV is recommended at the time of admission to animal shelters and many use a modified live virus (MLV) vaccine. From a diagnostic standpoint for dogs with suspected CDV infection, this is problematic because highly sensitive diagnostic real-time reverse transcription polymerase chain reaction (RT-PCR) tests are able to detect MLV virus in clinical samples. Real-time PCR can be used to quantitate amount of virus shedding and can differentiate vaccine strains from wild-type strains when shedding is high. However, differentiation by quantitation is not possible in vaccinated animals during acute infection, when shedding is low and could be mistaken for low level vaccine virus shedding. While there are gel-based RT-PCR assays for differentiation of vaccine strains from field strains based on sequence differences, the sensitivity of these assays is unable to match that of the real-time RT-PCR assay currently used in the authors' laboratory. Therefore, a real-time RT-PCR assay was developed that detects CDV MLV vaccine strains and distinguishes them from wild-type strains based on nucleotide sequence differences, rather than the amount of viral RNA in the sample. The test is highly sensitive, with detection of as few as 5 virus genomic copies (corresponding to 10 −1 TCID 50 ). Sequencing of the DNA real-time products also allows phylogenetic differentiation of the wild-type strains. This test will aid diagnosis during outbreaks of CDV in recently vaccinated animals.

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Section: Introductionmentioning

confidence: 99%

Real-time reverse transcription polymerase chain reaction method for detection of Canine distemper virus modified live vaccine shedding for differentiation from infection with wild-type strains

et al. 2014

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“…The System A detected RNA down to 1.1 x 10 RNA copiesµL -1 and was more sensitive than twostep probe based chemistry using the same primer pair (ELIA et al, 2006). Benefits of SYBR Green I One-…”

Section: Resultsmentioning

confidence: 99%

“…RT-qPCR assays have been applied to detect and quantify canine distemper virus in clinical specimens of naturally infected dogs or to distinguish vaccine strains from wild strains, using two-step and real-time probes chemistry (ELIA et al, 2006;SCAGLIARINI et al, 2007;FISCHER et al, 2013;WILkES et al, 2014). Recently, surface plasmon resonance (SPR), electrochemical impedance spectroscopy (EIS) and conjugated gold nanoparticles methodologies were also developed for CDV detection (BASSO et al, 2013;2015a;2015b).…”

Section: Introductionmentioning

confidence: 99%

“…Various samples types, such as blood, conjunctival swabs and urine, have been used for CDV diagnosis. Urine is useful in the ante mortem diagnosis of distemper, easier to collect than other body fluids and shows great sensitivity in different clinical presentations of canine distemper (GEBARA et al, 2004;ELIA et al, 2006). The aim of this study was to evaluate three commercial kits of One-Step RT-qPCR and to compare the analytical sensitivity of One-Step RT-qPCR (System A) and One-Step RT-qPCR combined with NESTED-qPCR (System B) to detect CDV RNA in urine samples from dogs with clinical suspicion of CDV infection.…”

Section: Introductionmentioning

confidence: 99%

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Canine distemper virus detection by different methods of One-Step RT-qPCR

et al. 2016

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Three commercial kits of One-Step RT-qPCR were evaluated for the molecular diagnosis of Canine Distemper Virus. Using the kit that showed better performance, two systems of Realtime RT-PCR (RT-qPCR) assays were tested and compared for analytical sensitivity to Canine Distemper RESUMO Três kits comerciais de One-Step RT-qPCR foram avaliados para o diagnóstico molecular do Vírus da Cinomose Canina.Utilizando o kit que apresentou melhor desempenho, dois sistemas de RT-PCR em tempo real (RT-qPCR) foram comparados quanto à sensibilidade analítica na detecção do RNA do Vírus da Cinomose Canina:One-Step RT-qPCR (Sistema A) e One-Step RT-qPCR seguido da NESTED-qPCR (Sistema B).Os limites de detecção dos dois sistemas foram determinados utilizando diluição

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“…This gene and its protein are currently subject to many studies due to its importance on transcription, replication and encapsidation of the RNA genome (Stettler and Zurbriggen 1995). In the same way, the sequencing of N gene is essential due to its use for molecular diagnostic, mainly in RT-PCR (Shin et al 1995, Frisk et al 1999, Kim et al 2001, Shin et al 2004 and Real Time RT-PCR (Elia et al 2006, Scagliarini et al 2007.…”

Section: Discussionmentioning

confidence: 99%

Identification of new genovariants of canine distemper virus in dogs from the State of Mexico by analyzing the nucleocapsid gene

2012

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RESUMENA nivel mundial, el distemper canino es una de las más importantes enfermedades virales en los perros, debido a su alta mortalidad y morbilidad. Es causada por el virus del distemper canino, un virus RNA, el cual ha demostrado que posee una elevada diversidad genética. En México no existen datos de epidemiología molecular sobre este virus, pero recientemente se ha registrado la presencia de una variante genética aparentemente exclusiva del país. Para determinar la diversidad genética del virus del distemper canino obtenido en el Estado de México, México, en este trabajo se analizaron muestras de perros que presentaban signos de la enfermedad. Para esto, las muestras obtenidas fueron procesadas por RT-PCR y la secuencia nucleotídica de un fragmento del gen N fue obtenida. El análisis de los datos fue realizado mediante filogenia molecular. Los resultados muestran que las secuencias del gen N pertenecen a siete genovariantes del virus del distemper canino que no habían sido previamente reportadas y se encuentran circulando en el Estado de México.

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Detection of canine distemper virus in dogs by real-time RT-PCR

Cited by 191 publications

References 14 publications

Real-time reverse transcription polymerase chain reaction method for detection of Canine distemper virus modified live vaccine shedding for differentiation from infection with wild-type strains

Real-time reverse transcription polymerase chain reaction method for detection of Canine distemper virus modified live vaccine shedding for differentiation from infection with wild-type strains

Canine distemper virus detection by different methods of One-Step RT-qPCR

Identification of new genovariants of canine distemper virus in dogs from the State of Mexico by analyzing the nucleocapsid gene

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