Avaliação molecular parcial do gene da hemaglutinina do vírus da cinomose canina

AUTOR(ES)

Fábio Juliano Negrão

FONTE

IBICT - Instituto Brasileiro de Informação em Ciência e Tecnologia

DATA DE PUBLICAÇÃO

27/02/2004

RESUMO

Urine and leucocytes from 56 dogs with canine distemper clinical diagnosis were evaluated by use in reverse transcription-polymerase chain reaction (RT-PCR) to canine distemper virus (CDV) hemagglutinin gene. In accordance with clinical signs the dogs was disposed in tree groups (A/systemic=17; B/neurological=8; C/systemic and neurological=31) the partial amplification from 721 base pare was possible 45 of 56 dogs. In 34 of 56 dogs the CDV was detected such in urine as in leucocytes. In 11 of 45 dogs only one biological sample was positive (urine=1 dog; leucocytes=10 dogs). It is advisable at least two biological sample to detect the CDV. The variability of canine distemper sings and the material choice may be to generate false-negative results principally en dogs with only systemic clinical sings (A group) where in 7 of 17 dogs was negative when compared with 1/8 B group and 3/31 C group, it is advisable at least two biological sample to detect the CDV. In other experiment 27 dogs positive and 5 negative by RT-PCR to CDV nucleoprotein gene were disposed in four groups in accordance with clinical sings (systemic=10 dogs; neurological=8 dogs; systemic and neurological= 9; control=5). To restriction fragment length polymorphism assay (RFLP) the enzymes Hinf I and Rsa I were selected to determine the RFLP pattern from 721 base pare (bp) fragment of haemagglutinin gene was amplified by using directly biological sample and the laboratory CDV strain Onderstepoor, Rockborn and Snyder Hill. Everyone PCR products after digestion with Hinf I were cut in two seeing fragment of 320 and 230 bp. The wild-type CDV PCR products later than digestion with Rsa I were cut into two seeing fragment around 360 and 230 bp that differ from the laboratory CDV strain. The Onderstepoort PCR product after than digest with Rsa I was cut in 363 and 327 bp. The RFLP pattern with Rsa I from Snyder Hill and Rockborn were the same two fragments with 363 and 358 bp. The Rockborn strain don’t have the genome in public database, but the computational analysis using public nucleotide sequence database and this study show the same RFLP pattern for Onderstepoort and Snyder Hill. The PCR and RFLP products were detected by electrophoresis in 2% agarose gels with ethidium bromide staining and the little fragments don’t be seeing. The molecular relationships showed in RFLP analysis suggest that wild-type current in north Paraná are unlike from laboratory CDV strains, and this different RFLP pattern may be characterize molecular distinction.

ASSUNTO(S)

cinomose canina virologia veterinária sanidade animal biologia molecular hemaglutinina canine distemper veterinary virology hemagglutinin

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