Porownanie klasycznej metody PCR oraz Real-Time Sybr-Green HRM PCR w rozpoznawaniu zakazen parwowirusowych u psow

• Autorzy: Adaszek L , Twarog D. , Zietek J. , Wojciechowski M. , Winiarczyk S.

Warianty tytułu

EN

Comparison of standard PCR and Real-Time PCR in the diagnose of canine parvoviral infections

• Języki publikacji: PL

Abstrakty

EN

The aim of this study was to compare a standard PCR and Sybr-Green HRM PCR in the diagnosis of canine parvoviral infections. A total of 22 feces samples were collected from dogs suspected of parvovirosis. The entire DNA for standard PCR and Real-Time PCR was isolated from the feces. In both methods this same pair of primers that allow the amplification of a fragment of VP 2 gene with a length of 1278 bp were used. The specificity of the obtained PCR products in the classical method were established based on the results of sequencing 8 out of 22 DNA probes and based on the comparison of their sequences with a CPV VP2 FJ 222823 sequence taken from the GeneBank. The specificity of Real-Time PCR products were established based on the analysis of their melting curve. In both standard and Real-Time PCR CPV DNA was detected in all 22 feces samples. The length of the obtained products was 1190 bp. To obtain a positive result in Real-Time PCR it was required to increase the number of the cycles from 30 to 60. The Ct values were between 43-53, and the analysis of the melting curve revealed that the Tm of Real-Time PCR products ranged between 80.5-85°C. Despite the results of this study indicating that both of these techniques are specific, sensitive, and repeated methods for detection of the CPV DNA, to shorten the time of Real-Time PCR the application of appropriate primers is required, which enables the amplification of shorter fragments of the DNA than those obtained in our study.

Słowa kluczowe

PL

psy; choroby zwierzat; parwowiroza psow; parwowirus psow; diagnostyka laboratoryjna; lancuchowa reakcja polimerazy; metoda Real Time PCR HRM; porownanie

• Wydawca: -
• Czasopismo: Medycyna Weterynaryjna
• Rocznik: 2009
• Tom: 65
• Numer: 10
• Opis fizyczny: s.683-686,fot.,rys.,bibliogr.

Twórcy

autor

Adaszek L

• Uniwersytet Przyrodniczy w Lublinie, ul.Gleboka 30, 20-612 Lublin

autor

Twarog D.

autor

Zietek J.

autor

Wojciechowski M.

autor

Winiarczyk S.

Bibliografia

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