A Loop-Mediated Isothermal Amplification Assay for Rapid Detection of Cyprinid Herpesvirus 2 in Gibel Carp (Carassius auratus gibelio)

作者：Zhang, Hui; Zeng, Lingbing^*; Fan, Yuding; Zhou, Yong; Xu, Jin; Ma, Jie

来源：The Scientific World Journal, 2014, 2014: 716413.

DOI：10.1155/2014/716413

• HEMATOPOIETIC NECROSIS VIRUS
• SPOT SYNDROME VIRUS
• SENSITIVE DETECTION
• KOI-HERPESVIRUS
• GOLDFISH
• IRIDOVIRUS
• SHRIMP
• FISH
• L.

A rapid and sensitive loop-mediated isothermal amplification (LAMP) assay for Cyprinid herpesvirus 2 (CyHV-2) detection in gibel carp was developed. Following cloning and sequencing of the putative DNA helicase gene of CyHV-2 isolate from China, a set of four specific primers was designed based on the sequence. The MgCl2 concentration and the reaction temperature were optimized to 6 mM, 64 degrees C, respectively. LAMP products were detected by visual inspection of a color change due to addition of SYBR Green I stain. The specificity and sensitivity of the LAMP assay were determined. No cross-reaction was observed with other fish DNA viruses including eel herpesvirus, koi herpesvirus, and Chinese giant salamander iridovirus. The LAMP assay was found to be equally sensitive as nested PCR. A comparative evaluation of 10 fish samples using LAMP and nested PCR assays showed an overall correlation in positive and negative results for CyHV-2. These results indicate that the LAMP assay is simple, sensitive, and specific and has a great potential use for CyHV-2 detection in the laboratory and field.

• 出版日期2014
• 单位中国水产科学研究院; 中国水产科学研究院长江水产研究所