Comparison of one‐step RT‐PCR and a nested PCR for the detection of canine distemper virus in clinical samples

Shin, Young-Hak; Cho, K O; Cho, H S; Kang, Sang Kyu; Kim, H J; Kim, Y H; Park, H S; Park, N Y

doi:10.1111/j.1751-0813.2004.tb14651.x

Cited by 53 publications

(69 citation statements)

References 14 publications

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“…All three viruses are present in South Korea, where 5.1% of dogs are seropositive for CIV and 12.8% are seropositive for CRCoV [2,17]. Using one step RT-PCR, CDV has been detected in 21-70% of clinical samples such as blood, urine, nasal swabs, and saliva [3,23].…”

mentioning

confidence: 99%

“…CRCoV replicates in the upper respiratory tract and can, therefore, be detected in respiratory tissues and nasal swabs [19]. CDV can be detected in various body fluids including urine, blood, saliva, nasal secretions, and cerebrospinal fluid [15,23]. Because all these three viruses can be detected in nasal secretions, hence nasal swabs were used for the mRT-PCR assay.…”

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confidence: 99%

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Simultaneous Detection of Canine Respiratory Disease Associated Viruses by a Multiplex Reverse Transcription-Polymerase Chain Reaction Assay

Jeoung

Song

Jeong

et al. 2013

The Journal of Veterinary Medical Science

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ABSTRACT. A multiplex reverse transcription polymerase chain reaction (mRT-PCR) assay was developed for the simultaneous detection of canine distemper virus (CDV), canine respiratory coronavirus (CRCoV) and canine influenza virus (CIV). These viral pathogens are all causative agents of canine infectious respiratory disease (CIRD). The sensitivity and specificity of the mRT-PCR were determined by comparing it to a rapid antigen test (RAT) or immuno-chromatography test kit and reverse transcription-polymerase chain reaction (RT-PCR) in the detection of CDV, CRCoV and CIV antigens present in 100 clinical samples (nasal swabs and whole blood samples) from 50 dogs with respiratory disease symptoms. This study revealed that mRT-PCR had almost exactly the same performance or results were almost 100% in agreement with that of RT-PCR and RAT both in terms of the assay sensitivity and specificity which was more highly evident in detecting CIV, CDV and CRCoV antigens present in canine nasal swab samples. Therefore, this assay could be a better alternative for the definitive and simultaneous ante-mortem detection of the three viral pathogens that cause CIRD by using nasal swabs.

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confidence: 99%

mentioning

confidence: 99%

Simultaneous Detection of Canine Respiratory Disease Associated Viruses by a Multiplex Reverse Transcription-Polymerase Chain Reaction Assay

Jeoung

Song

Jeong

et al. 2013

The Journal of Veterinary Medical Science

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“…When using conventional reaction, the limit of detection of N gene of CDV was increased hundred fold using the One-Step RT-PCR combined with the NESTED PCR (kIM et al, 2001). The CDV was detected in 50.0% of urine samples from symptomatic dogs by One-Step RT-PCR and in 75.0% of urine samples when combined with NESTED-PCR (SHIN et al, 2004). Thus, in the present study, the System A showed superior analytical sensitivity compared to System B due to greater reaction efficiency.…”

Section: Resultsmentioning

confidence: 58%

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%

“…RNA was eluted in 15 µl of nuclease free water (Fermentas, USA). A pair of primers previously reported by (Shin et al 2004) was used to amplify a 297 bp fragment of the N gene. RT-PCR was performed using the Access Quick RT-PCR Kit (Promega, USA).…”

Section: Methodsmentioning

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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Comparison of one‐step RT‐PCR and a nested PCR for the detection of canine distemper virus in clinical samples

Abstract: The nested PCR detected CDV in blood, urine, nasal swab and saliva more frequently than did the one-step RT-PCR. Therefore, this assay should be a useful aid to antemortem diagnosis of CDV infections in dogs.

Cited by 53 publications

References 14 publications

Simultaneous Detection of Canine Respiratory Disease Associated Viruses by a Multiplex Reverse Transcription-Polymerase Chain Reaction Assay

Simultaneous Detection of Canine Respiratory Disease Associated Viruses by a Multiplex Reverse Transcription-Polymerase Chain Reaction Assay

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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