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Rapid visual detection of cyprinid herpesvirus 2 by recombinase polymerase amplification combined with a lateral flow dipstick.
- Source :
-
Journal of Fish Diseases . Aug2018, Vol. 41 Issue 8, p1201-1206. 6p. - Publication Year :
- 2018
-
Abstract
- Herpesviral haematopoietic necrosis (HVHN), caused by cyprinid herpesvirus 2 (CyHV-2), causes significant losses in crucian carp (Carassius carassius) aquaculture. Rapid and convenient DNA assay detection of CyHV-2 is useful for field diagnosis. Recombinase polymerase amplification (RPA) is a novel isothermal DNA amplification and detection technology that can amplify DNA within 30 min at ~37°C by simulating in vivo DNA recombination. Herein, a rapid and convenient detection assay based on RPA with a lateral flow dipstick (LFD) was developed for detecting CyHV-2. The highly conserved ORF72 of CyHV-2 was targeted by specific and sensitive primers and probes. The optimized assay takes only 15 min at 38°C using a water bath, with analysis of products by 2% agarose gel electrophoresis within 30 min. A simple lateral flow strip based on the unique probe in reaction buffer was developed for visualization. The entire RPA-LFD assay takes 50 min less than the routine PCR method, is 100 times more sensitive and displays no cross-reaction with other aquatic viruses. The combined isothermal RPA and lateral flow assay (RPA-LFD) provides a simple, rapid, reliable method that could improve field diagnosis of CyHV-2 when resources are limited. [ABSTRACT FROM AUTHOR]
- Subjects :
- *CYPRINIDAE
*HERPESVIRUSES
*POLYMERASES
*AQUACULTURE
*GENE amplification
Subjects
Details
- Language :
- English
- ISSN :
- 01407775
- Volume :
- 41
- Issue :
- 8
- Database :
- Academic Search Index
- Journal :
- Journal of Fish Diseases
- Publication Type :
- Academic Journal
- Accession number :
- 130923533
- Full Text :
- https://doi.org/10.1111/jfd.12808