Polymerase chain reaction amplification of genomic fragments of bovine herpesvirus-1

Abstract

Especial conditions were developed for the amplification of five DNA segments from US region of BHV-1 by polymerase chain reaction. In order to eliminate most nonspecific products it was found that addition of three cosolvents DMSO, glycerol and NP 40 was a simple method for increasing the specificity of amplification.

Development of a Polymerase Chain Reaction and Restriction Typing Assay for the Diagnosis of Bovine Herpesvirus 1, Bovine Herpesvirus 2, and Bovine Herpesvirus 4 Infections

“…Several molecular diagnostic assays based on the polymerase chain reaction (PCR) have been described, 1,2,4,16,17,20,22 but in most cases these assays are employed to detect a specific herpesvirus 2,4,10,20,22 or a group of related herpesviruses. 1,16,17 A multiplex PCR method coupled with endonuclease digestion for restriction fragment length polymorphism (PCR-RFLP) analysis of the amplicons is described here.…”

Padronização e comparação de técnicas de reação em cadeia por polimerase (PCR) para detecção do metapneumovírus humano em secreções respiratórias

“…Diversos trabalhos comprovaram o aumento da especificidade na técnica de PCR causada pela adição de co-solventes, entre os quais o glicerol (Navaneetham et al, 1998;Paragas et al, 1998;Cândido et al, 2000) o dimetilsufóxido (Paragas et al, 1998; Whinship, 1989;Bachmann et al, 1990;Cândido et al, 2000;Chakrabarti et al, 2001) e a formamida (Sarkar et al, 1990;Paragas et al, 1998). .…”

“…Se analisarmos os resultados das técnicas separadamente, verificou-se que a técnica em tempo real detectou 11 amostras positivas em 228 (4,8%) -tabela 6. (Navaneetham et al, 1998;Paragas et al, 1998;Cândido et al, 2000).…”