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2011 | 67 | 07 |

Tytuł artykułu

Zastosowanie real time RT-PCR do wykrywania wirusa biegunki bydła i choroby błon śluzowych (BVD-MD) typu 1 i typu 3 w materiale biologicznym

Warianty tytułu

EN
Application of real time RT-PCR for the detection of type 1 and type 3 of bovine viral diarrhea virus (BVDV) in biological material

Języki publikacji

PL

Abstrakty

EN
The aim of the study was to use real time RT-PCR for the detection of genetic material of bovine viral diarrhea virus (BVDV) type 1 and type 3 in serum and milk samples. Material tested included the fetal calf serum used for cell culture, serum samples from healthy calves and from calves experimentally inoculated with BVDV type 1 and type 3, and milk samples (pasteurized and treated with ultra high temperature). Sensitivity of the test was 200 copies of RNA per reaction (10⁵ viral RNA copies per ml) for both types of BVDV, using dedicated primers and standards. Detection limit was 10² tissue culture infectious dose 50 (TCID₅₀) and 1 TCID₅₀ for type 1 and type 3, respectively. Diagnostic specificity of the method was 100%. Out of 10 samples of milk, 3 were positive for BVDV type 1, while none was positive for type 3. On the other hand, BVDV type 3 was found in 6 out of 10 samples of fetal calf serum. Real Time RT-PCR for BVDV type 1 and type 3 proved to be a highly sensitive and highly specific technique, enabling the detection of viral genetic material in various samples, even when its detection by virus isolation or antigen ELISA tests is impossible because of virus inactivation by such processes as high temperature, gamma irradiation, or the presence of virus neutralizing antibodies.

Wydawca

-

Rocznik

Tom

67

Numer

07

Opis fizyczny

s.483-487,rys.,tab.,bibliogr.

Twórcy

autor
  • Department of Clinical Sciences, Swedish University of Agricultural Sciences (SLU), BOX 7054, SE 750 07 Uppsala, Szwecja
autor

Bibliografia

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  • 3.Baxi M., McRae D., Baxi S., Greiser-Wilke I., Vilcek S., Amoako K., Deregt D.: A one-step multiplex real-time RT-PCR for detection and typing of bovine viral diarrhea viruses. Vet. Microbiol. 2006, 116, 37-44.
  • 4.Decaro N., Mari V., Lucente M. S., Colainni M. L., Cirone F., Losurdo M., Cordioli P., Buonavoglia C.: Virus della diarrea bovina tipo 3 associato a malattia respiratoria. XII Congresso Nationale S.I.Di.L.V., Genewa 27-29. 10.2010, s. 96-97.
  • 5.Drew T. W., Napp F., Paton D. P.: The detection of bovine viral diarrhoea virus in bulk milk samples by the use of a single-tube RT-PCR. Vet. Microbiol. 1999, 64, 145-154.
  • 6.Frölich K., Jung S., Ludwig A., Lieckfeldt D., Gibert P., Gauthier D., Hars J.: Detection of a newly described pestivirus of Pyrenean chamois (Rupicapra pyrenaica pyrenaica) in France. J. Wildl. Dis. 2005, 41, 606-610.
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  • 9.Liu L., Xia H., Belák S., Baule C.: A TaqMan real-time RT-PCR assay for selective detection of atypical bovine pestiviruses in clinical samples and biological products. J. Virol. Methods 2008, 154, 82-85.
  • 10.Makoschey B., v. Gelder P. T. J. A., Keijsers V., Goovaerts D.: Bovine viral diarrhoea virus antigen in foetal calf serum batches and consequences of such contamination for vaccine production. Biologicals 2003, 31, 203-208.
  • 11.Pejsak Z., Truszczyński M.: Zespół zapalenia mięśnia sercowego prosiąt ssących - nowa wirusowa choroba świń. Życie Wet. 2007, 82, 734-736.
  • 12.Pellerin C., van den Hurk J., Lecomte J., Tijssen P.: Identification of a new group of bovine viral diarrhea virus strains associated with severe outbreaks and high mortalities. Virology 1994, 203, 260-268.
  • 13.Polak M. P., Rola J., Żmudziński J. F.: Contamination of fetal bovine serum with bovine viral diarrhoea virus. Bull. Vet. Inst. Pulawy 2008, 52, 501-505.
  • 14.Polak M. P., Zmudzinski J. F.: Prevalence of bovine viral diarrhea virus infection in bulls in artificial insemination centers in Poland. Vet. Microbiol. 1999, 64, 253-257.
  • 15.Polak M. P., Żmudziński J. F.: Genotypowanie krajowych izolatów wirusa BVD-MD. Medycyna Wet. 2002, 58, 913-1004.
  • 16.Renshaw R. W., Ray R., Dubovi E. J.: Comparison of virus isolation and reverse transcription polymerase chain reaction assay for detection of bovine viral diarrhea virus in bulk milk tank samples. J. Vet. Diagn. Invest. 2000, 12, 184-186.
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  • 18.Stadejek T., Podgórska K., Pejsak Z.: Zastosowanie Real Time PCR do wykrywania zakażeń pestiwirusowych. Medycyna Wet. 2006, 62, 165-169.
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Typ dokumentu

Bibliografia

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Identyfikator YADDA

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