Application of N-PCR for diagnosis of distemper in dogs and fur animals

Rzeżutka, Artur; Mizak, B.

doi:10.1016/s0378-1135(02)00097-4

Cited by 33 publications

(19 citation statements)

References 9 publications

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“…The sensitivity of RT-PCR alone was inadequate enough to detect viral RNA. This work co related with the superiority of nested PCR as a RNA detection tool by Rzezutka et al, 2002 and falls in line to the higher sensitivity of detection as stated as Yong-Hwan Kim et al, 2001. Non-specific reactions were common in conventional antigen detection methods such as AGPT and DOT-ELISA.…”

Section: Nested Pcr Combined With Rt Pcrmentioning

confidence: 59%

Detection Of Canine Distemper Virus Using Nested PCR

Muthiah¹,

Parthiban²

2008

Int. Jour. Appl. Bio-Eng.

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Canine Distemper Virus is one of the most contagious RNA virus, which causes many fatal respiratory problems in dogs, seals, Red foxes, lions, badgers etc.,. Detection of CDV poses a very big and important task among veterinarians. This report mainly emphasis on the isolation and detection of the canine distemper virus in a novel way. Nasal and ocular samples from 25 dogs were collected from the clinical ward. Out of that, the first 5 were cultured in cell lines, namely MDCK and VERO. These cultured samples were passaged each for about 7 times which was sufficient enough for PCR detection. Field samples and cultured samples were then processed. The Viral RNA was extracted from cultured samples and the corresponding cDNAs were synthesized. Finally RT-PCR was done and it was followed by nested PCR. Out of the 25 field samples tested, 2 of them were detected positive and out of the 5 cultured samples, 3 were detected positive. Thus, the sensitivity of detection is shown to increase by about 40% when cultured, as the RNA content is prone to increase in cell culture.ergillus niger, Surface culture, Banana peel, Biomass, Fermentation.

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Section: Nested Pcr Combined With Rt Pcrmentioning

confidence: 59%

Detection Of Canine Distemper Virus Using Nested PCR

Muthiah¹,

Parthiban²

2008

Int. Jour. Appl. Bio-Eng.

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“…The application of molecular techniques (for example, gel-based RT-PCR) in CDV diagnosis has been successfully used in different types of samples collected from animals clinically suspected of infection (Frisk et al 1999, Kim et al 2001, Rzezutka & Mizak 2002, Gebara et al 2004. It is a specific, practical and rapid tool in CDV diagnosis ante mortem considering the non-specific clinical signs of the disease, but it is not a quantitative and very sensitive assay.…”

Section: Discussionmentioning

confidence: 99%

“…ELISA assay may detect high antibody titers to CDV for several months after vaccination or after subclinical or clinical infection, turning false positive results (Martella et al 2008). Molecular assays, such as RT-PCR (Frisk et al 1999, Kim et al 2001, Rzezutka & Mizak 2002, Gebara et al 2004) are sensitive and specific, but not quantitative.…”

Section: Canine Distemper Virus Detection In Asymptomatic Andmentioning

confidence: 99%

Canine distemper virus detection in asymptomatic and non vaccinated dogs

et al. 2010

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Samples from eleven domestic dogs with no signs of canine distemper and not vaccinated at the month of collection were used. Canine distemper virus vaccine samples in VERO cells were used as positive controls. RNA was isolated with Trizol®, and treated with a TURBO DNA-free kit. Primers were designed for canine distemper virus nucleocapsid protein coding region fragment amplification (84 bp). Canine β-actin (93 bp) was utilized as the endogenous control for normalization. Quantitative results of real time PCR generated by ABI Prism 7000 SDS Software showed that 54.5% of dogs with asymptomatic canine distemper were positive for canine distemper virus. Dissociation curves confirmed the specificity of the real time PCR fragments. This technique could detect even a few copies of viral RNA and identificate subclinically infected dogs providing accurate diagnosis of this disease at an early stage.

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“…CDV immunoassays for detection of CDV antigen, such as immunofluorescence staining and ELISA [28][29][30][31][32], in general are not sensitive enough to detect CDV in asymptomatic dogs or during acute CD infection [31]. Various conventional, nested, and real-time reverse transcription polymerase chain reaction (RT-PCR) assays, which offer high degrees of detection sensitivity and specificity, have also been developed for CDV [33][34][35][36][37][38][39][40][41][42]. However, requiring an experienced technician and a relatively expensive thermocycler, these assays have been applied to CDV diagnosis mostly in professional laboratories.…”

Section: Introductionmentioning

confidence: 99%

Rapid and sensitive detection of canine distemper virus by one-tube reverse transcription-insulated isothermal polymerase chain reaction

et al. 2014

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Background: Canine distemper virus (CDV) has been associated with outbreaks of canine infectious respiratory disease in shelters and boarding kennel environments. POCKIT TM Nucleic Acid Analyzer is a field-deployable device capable of generating automatically interpreted insulated isothermal polymerase chain reaction (iiPCR) results from extracted nucleic acid within one hour. In this study, reverse transcription iiPCR (RT-iiPCR) was developed to facilitate point-of-need diagnosis of CDV infection. Results: Analytical sensitivity (limit of detection 95%) of the established CDV RT-iiPCR was about 11 copies of in vitro transcribed RNA per reaction. CDV RT-iiPCR generated positive signals from CDV, but not Bordetella bronchiseptica, canine parvovirus, canine herpesvirus, canine adenovirus 2, canine influenza virus (subtype H3N8), canine parainfluenza virus, and canine respiratory coronavirus. To evaluate accuracy of the established reaction in canine distemper clinical diagnosis, 110 specimens from dogs, raccoons, and foxes suspected with CDV infection were tested simultaneously by CDV RT-iiPCR and real-time RT-PCR. CDV RT-iiPCR demonstrated excellent sensitivity (100%) and specificity (100%), compared to real-time RT-PCR.

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Application of N-PCR for diagnosis of distemper in dogs and fur animals

Cited by 33 publications

References 9 publications

Detection Of Canine Distemper Virus Using Nested PCR

Detection Of Canine Distemper Virus Using Nested PCR

Canine distemper virus detection in asymptomatic and non vaccinated dogs

Rapid and sensitive detection of canine distemper virus by one-tube reverse transcription-insulated isothermal polymerase chain reaction

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