The development of a loop-mediated isothermal amplification assay for rapid and sensitive detection of abalone herpesvirus DNA.
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 60min, 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.
['Chen|M H|MH|', 'Kuo|S T|ST|', 'Renault|T|T|', 'Chang|P H|PH|']
D000818:Animals D017931:DNA Primers D004279:DNA, Viral D049851:Gastropoda D006564:Herpesviridae D021141:Nucleic Acid Amplification Techniques D012680:Sensitivity and Specificity D013696:Temperature
Loop-mediated isothermal amplification Abalone Herpesvirus SYBR green PCR Haliotis diversicolor supertexta Polymerase chain reaction