Naslov
Differentiation of animal virus strains by restriction endonuclease analysis

Autori
Lojkić, Ivana ; Biđin, Zdenko ; Grce, Magdalena ; Husnjak, Koraljka ; Pokrić, Biserka

Vrsta, podvrsta i kategorija rada
Sažeci sa skupova, sažetak, znanstveni

Izvornik
2nd scientific symposium with international participation - 45 years of molecular biology in Croatia and 50 years of double helix / Ambirović Ristov, Andreja ; Brozović, Anamaria - Zagreb, 2003

Skup
45 years of molecular biology in Croatia and 50 years of double helix

Mjesto i datum
Zagreb, Hrvatska, 20.11.2003. - 21.11.2003

Vrsta sudjelovanja
Poster

Vrsta recenzije
Nije recenziran

Ključne riječi
virus zarazne bolesti burze; RT-PCR; restrikcijske endonukleaze
(infectious bursal disease virus; RT-PCR; restriction endonuclease)

Sažetak
The restriction enzyme (RE) analysis of the reverse transcription (RT)/polymerase chain reaction (PCR) products of viral genome segments is proposed as a method for the differentiation of the field isolates of animal viruses. The accuracy of the method was tested on the infectious bursal disease viruses (IBDV) responsible for the outbreak of the disease among chicken in Croatia during a 5 year period. The analysis was conducted with the 422 bp hypervariable region of the VP2 gene. The RT/PCR products were digested using the restriction enzymes CfoI, SacI, SspI, StuI and TaqI. The knowledge of the nucleotide sequence recognized by the endonuclease, the literature and the GenBank nucleotide data for the analyzed VP2 gene region, as well as the size of the fragments obtained by the endonuclease digestion, enabled the determination of the nucleotide position corresponding to a restriction site. The differentiation of the analysed IBDV species was obtained by comparing the position of the restriction sites in analysed IBDV species with the data reported elsewhere. The RE-RT/PCR analyses showed that 98% of IBDV positive tissue samples were infected with very virulent IBDV strains. This result agreed and explained a high rate of chicken morbidity and mortality in the period of sample collection.

Izvorni jezik
Engleski

Znanstvena područja
Biologija, Veterinarska medicina

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