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Development of a polymerase chain reaction for the detection of abalone herpesvirus infection based on the DNA polymerase gene ArchiMer
Chen, M.h.; Kuo, S. T.; Renault, Tristan; Friedman, C. S.; Chang, P. H..
A 5781-base pair (bp) fragment of genomic DNA from the Taiwanese abalone herpesvirus was obtained and showed 99% (5767/5779) homology in the nucleotide sequence and 99% (1923/1926) in the amino acid sequence with the DNA polymerase gene of the abalone herpesvirus strain Victoria/AUS/2007. Homology of the amino acid sequence with the DNA polymerase of ostreid herpesvirus 1 was 30% (563/1856). In this study, a PCR-based procedure for detecting herpesvirus infection of abalone, Haliotis diversicolor supertexta, in Taiwan was developed. The method employed primer sets targeting the viral DNA polymerase gene, and was able to amplify DNA fragments of the expected size from infected samples. Primer sets of 40f and 146r were designed for amplification of an...
Tipo: Text Palavras-chave: Herpes virus; Abalone; Haliotis diversicolor supertexta; DNA polymerase; Polymerase chain reaction.
Ano: 2012 URL: http://archimer.ifremer.fr/doc/00098/20952/19675.pdf
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The development of a loop-mediated isothermal amplification assay for rapid and sensitive detection of abalone herpesvirus DNA ArchiMer
Chen, Meilin; Kuo, S. T.; Renault, Tristan; Chang, P. H..
A loop-mediated isothermal amplification (LAMP) assay was developed for the detection of abalone herpesvirus DNA. Two pairs of primers were designed, based on the sequence of the DNA polymerase gene of abalone herpesvirus. The reaction temperature and time were optimized to 63 °C and 60 min, respectively. LAMP amplicons were analyzed by 2% agarose gel electrophoresis or by visual inspection of a colour change emitted by fluorescent dye. The method developed was specific for the detection of abalone herpesvirus, without cross-reactions with other tested herpesviruses including ostreid herpesvirus 1 (OsHV-1), European eel herpesvirus, koi herpesvirus (KHV) and an avian herpesvirus. The LAMP assay was 100 folds more sensitive than a conventional PCR and 10...
Tipo: Text Palavras-chave: Herpesvirus; Abalone; Haliotis diversicolor supertexta; Polymerase chain reaction; Loop-mediated isothermal amplification; SYBR green PCR.
Ano: 2014 URL: https://archimer.ifremer.fr/doc/00166/27690/25882.pdf
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