Author/Editor		Zorman-Rojs, O; Krapež, U; Grom, J; Barlič-Maganja, D
Title		Molecular detection and pathotyping of paramyxovirus type 1 isolates (Newcastle disease virus)
Translated title		Molekularno dokazovanje in tipizacija patogenosti izolatov paramiksovirusov tipa 1 (virusa atipične kokošje kuge)
Type		članek
Source		Slov Vet Res
Vol. and No.		Letnik 39, št. 1
Publication year		2002
Volume		str. 39-45
Language		eng, slo
Abstract		An RT-PCR method for the detection of paramyxovirus type 1 (PMV-1) Newcastle disease virus - of different pathotypes is described. Degenerate oligonucleotide primers, selected from the fusion protein coding gene, amplified the fragments of 254 bp. The nucleotide sequences of the obtained RT-PCR products were determined by direct nucleotide sequencing. Different reference PMV-1 strains and four PMV-1 isolates, one of chicken and three of pigeon origine, were included in the investigation. The amino acid sequences in the fusion protein cleavage site were deduced and the pathotype of the Slovene PMV-1 isolates was predicted. The chicken isolate had the fusion protein cleavage sequence (112R-R-Q-K-R116) of most known velogenic Newcastle disease virus (NDV) strains. The amino acid sequences, from the fusion protein cleavage site of the pigeons PMV-1 (112G-R-Q-K-R-F117), grouped these isolates between mesogenic strains. The RT-PCR detection method described, coupled with direct nucleotide sequencing, can be used for the diagnosis of Newcastle disease, its epidemiology and for predicting the pathotype of an NDV isolate.
Summary		Opisali smo dokazovanje nukleinske kisline paramiksovirusa tip 1 (PMV-1) virusa atipične kokošje kuge - z enostopenjsko metodo reverzne transkripcije s polimerazno verižno reakcijo (RT-PCR). Izbrali smo začetne oligonukleotide za pomnoževanje gena, ki kodira virusni fuzijski protein, in dobili produkt velikosti 254 bp. Z metodo direktnega sekveniranja smo določili nukleotidno zaporedje dobljenega produkta. V preiskavo smo vključili različne referenčne seve PMV-1 in štiri lastne izolate PMV-1; eden je bil izoliran pri piščancih, trije pa pri golobih. Na podlagi aminokislinske sekvence cepitvenega mesta fuzijskega proteina smo virusnim izolatom določili stopnjo patogenosti. Primerjava sekvenc aminokislin na mestu cepitve fuzijskega proteina je pokazala, da je virus izoliran iz piščancev (112-R-Q-K-R116), velogeni sev virusa atipične kokošje kuge, tri golobje izolate pa smo glede na sekvenco (112G-R-Q-K-R-F117) uvrstili med mesogene seve NDV. Opisana diagnostična metoda je primerna za dokazovanje virusa atipične kokošje kuge, za epidemiološke študije kot tudi za presojo patogenosti virusnih izolatov.
Descriptors		NEWCASTLE DISEASE VIRUS POLYMERASE CHAIN REACTION BIRDS AMINO ACID SEQUENCE