The development of a loop-mediated isothermal amplification assay for rapid and sensitive detection of abalone herpesvirus DNA
|Author(s)||Chen Meilin1, 3, Kuo S. T.2, Renault Tristan4, Chang P. H.1|
|Affiliation(s)||1 : Natl Taiwan Univ, Sch Vet Med, Inst Vet Med, Taipei 10764, Taiwan.
2 : Natl Inst Anim Hlth, Tansui, Taiwan.
3 : Tzu Chi Coll Technol, Hualien, Taiwan.
4 : IFREMER, Unite Sante Genet & Microbiol Mollusques, Lab Genet & Pathol Mollusques Marins, F-17390 La Tremblade, France.
|Source||Journal Of Virological Methods (0166-0934) (Elsevier Science Bv), 2014-02 , Vol. 197 , P. 199-203|
|WOS© Times Cited||8|
|Keyword(s)||Herpesvirus, Abalone, Haliotis diversicolor supertexta, Polymerase chain reaction, Loop-mediated isothermal amplification, SYBR green PCR|
|Abstract||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.|