A sensitive and specific hyperbranched rolling circle amplification assay and test strip for white spot syndrome virus

• Published in: Journal of AOAC International 2014-09, Vol.97 (5), p.1410-1415
• Main Authors: Zhao, Yu-Ran, Yin, Wei-Li, Yue, Zhi-Qin, Li, Ba-Fang
• Format: Article
• Language: English
• Subjects: Animals; Control; Diseases; Distribution; Nucleic Acid Amplification Techniques - methods; Pandalidae - virology; Prevention; Reagent Strips; Sensitivity and Specificity; Shrimps; Virus diseases; White spot syndrome virus 1 - genetics; White spot syndrome virus 1 - isolation & purification
• ISSN: 1060-3271; 1944-7922
• DOI: 10.5740/jaoacint.13-387

White spot syndrome virus (WSSV) is a global threat to the prawn industry, and there is no simple method for field-based testing of this virus. We designed a padlock probe and primers to the capsid protein gene VP28 of WSSV, and established a hyperbranched rolling circle amplification (HRCA) assay and a corresponding strip-based test. The assay and the test strip both had similar high accuracy and specificity, and their sensitivity was about 10 copies/μL, which is 100 times higher than conventional PCR. In this study, 68 batches of prawns were tested for WSSV with the HRCA assay and test strip, and the results were compared with the PCR assay. The results indicated that both the assay and test strip had accuracy similar to each other and to the PCR results. However, the assay and strip were more sensitive and user-friendly than PCR. Establishment of this method will provide a rapid detection of WSSV and also a basis for field-based detection of animal disease.