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Titolo:
AMPLIFICATION OF STRAINS OF BOVINE HERPESVIRUS-1 BY USE OF POLYMERASECHAIN-REACTION WITH PRIMERS IN THE THYMIDINE KINASE REGION
Autore:
KIBENGE FSB; HARRIS LM; MCKENNA PK; WADOWSKA D; YASON CV;
Indirizzi:
UNIV PRINCE EDWARD ISL,ATLANTIC VET COLL,DEPT PATHOL & MICROBIOL CHARLOTTETOWN C1A 4P3 PE CANADA
Titolo Testata:
American journal of veterinary research
fascicolo: 9, volume: 55, anno: 1994,
pagine: 1206 - 1212
SICI:
0002-9645(1994)55:9<1206:AOSOBH>2.0.ZU;2-3
Fonte:
ISI
Lingua:
ENG
Soggetto:
EQUINE HERPESVIRUS; DEFICIENT MUTANTS; VIRUS; PCR; TYPE-1; GENE; DNA; SEQUENCES; DIAGNOSIS; LOCATION;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Science Citation Index Expanded
Citazioni:
31
Recensione:
Indirizzi per estratti:
Citazione:
F.S.B. Kibenge et al., "AMPLIFICATION OF STRAINS OF BOVINE HERPESVIRUS-1 BY USE OF POLYMERASECHAIN-REACTION WITH PRIMERS IN THE THYMIDINE KINASE REGION", American journal of veterinary research, 55(9), 1994, pp. 1206-1212

Abstract

A primer pair was designed from the published nucleotide sequence of the coding region of the bovine herpesvirus 1 (BHV-1) thymidine kinase(tk) gene for use in detection of the virus by use of polymerase chain reaction (PCR) amplification of 12 BHV-1 strains (3 ATCC and 9 localisolates). A fk deletion mutant BHV-1, and 2 BHV-4 strains from ATCC were used as negative controls. One strain each of feline herpesvirus,equine herpesvirus, and bovine adenovirus, and 2 noninoculated bovinecultured cells-bovine fetal testis and Madin-Darby bovine kidney-alsowere examined to verify specificity of the primers. A PCR product, 183 bp long, was detected by ethidium bromide staining after agarose gelelectrophoresis, when purified DNA from cell cultures infected with BHV-1 strain LA was used as template. Specificity of the PCR product was confirmed by restriction digestion with Sac If enzyme and Southern blot hybridization. Amplification was detected by ethidium bromide staining of agarose gels and/or Southern blot hybridization with the radiolabeled PCR product of the LA strain in similarly prepared DNA templates of 5 other BHV-1 strains, 2 obtained from ATCC and 3 of the 9 localisolates. In a modified PCR protocol, using virus suspensions treatedwith a nucleic acid-releasing cocktail, substantial amplification wasobtained for the 3 BHV-1 strains from ATCC and for all 9 local bovineherpesvirus held isolates. A fk deletion mutant BHV-1 strain NG dltk,2 BHV-4 strains, a feline herpesvirus, an equine herpesvirus, and a bovine adenovirus obtained from ATCC, and noninoculated bovine fetal testis and Madin-Darby bovine kidney cell cultures amplified either frompurified DNA or from nucleic acid-releasing cocktail-treated suspensions had negative results of ethidium bromide staining and Southern blot hybridization. These data indicate that PCR amplification, using primers in the fk gene region, is valuable for detection of BHV-1 strainsgrown in cell culture.

ASDD Area Sistemi Dipartimentali e Documentali, Università di Bologna, Catalogo delle riviste ed altri periodici
Documento generato il 06/06/20 alle ore 07:34:19