期刊
JOURNAL OF VIROLOGICAL METHODS
卷 196, 期 -, 页码 199-203出版社
ELSEVIER SCIENCE BV
DOI: 10.1016/j.jviromet.2013.11.011
关键词
Herpesvirus; Abalone; Haliotis diversicolor supertexta; Polymerase chain reaction; Loop-mediated isothermal amplification; SYBR green PCR
资金
- Council of Agriculture, Taiwan
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 folds less sensitive than a SYBR Green PCR. These results indicate that the developed LAMP assay is a simple, rapid, sensitive, specific and reliable technique for the detection of abalone herpesvirus. (C) 2013 Elsevier B.V. All rights reserved.
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