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Detection and differentiation of waterfowl parvoviruses by PCR

100%

Wozniakowski G., Kozdrun W., Samorek-Salamonowicz E.

Bulletin of the Veterinary Institute in Puławy

2010

tom 54

nr 3

The presented study aimed at application of the PCR methods for the detection and differentiation of goose parvovirus (GPV) and Muscovy duck parvovirus (MDPV) in the liver, spleen, and faeces of infected geese and in litter from infected farms. For the improvement of specificity and efficiency of the methods, as well as for overcoming the frequent inhibition of PCR in samples extracted from faeces and litter, the "touchdown" thermal profile with additive of betaine were used. The isolation of the virus in goose embryo fibroblasts was used as a verifying method for GPV and MDPV detection. The presence of the cytopathic effect in infected cell cultures allowed for the detection of the both parvoviruses but not for their differentiation. As a result of this study, PCR methods for the fast detection of GPV and MDPV in field samples of visceral organs and faeces of infected geese and in their litter were developed.

Touchdown PCR for the detection of waterfowl parvoviruses

100%

Wozniakowski G., Kozdrun W., Samorek-Salamonowicz E., Krol K.

Bulletin of the Veterinary Institute in Puławy

2009

tom 53

nr 1

3-7

Sixteen field strains of goose parvovirus (GPV) and DNA extracted from standard strain of Muscovy duck parvovirus (MDPV) were used. PCR was used for the amplification of the VP3 structural protein encoding region. In the case of all GPV strains and one MDPV strain the presence of specific product about 1,604 bp long was observed. The essential conditions, which had the influence on the specificity, sensitivity, and efficiency of the amplification reaction, were optimised. In order to eliminate unspecific interactions between template and primers touchdown PCR was applied. Additionally, for specificity and efficiency improvement, betaine (N, N, N - trimethylglycine) was added. The conducted optimisation steps of PCR allowed for the identification of the both parvoviruses.

Ograniczanie wyników

2 Bulletin of the Veterinary Institute in Puławy

2 Kozdrun W.

2 Samorek-Salamonowicz E.

2 Wozniakowski G.

1 Krol K.

1 2010

1 2009

Wyniki wyszukiwania

Detection and differentiation of waterfowl parvoviruses by PCR

Touchdown PCR for the detection of waterfowl parvoviruses