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Add To Calendar 28/04/2016 11:00:0028/04/2016 11:20:00America/ChicagoAsian-Pacific Aquaculture 2016POCKITTM SYSTEM: A RAPID, SIMPLE, AND SENSITIVE ON-SITE DETECTION TOOL FOR Enterocytozoon hepatopenaei   Crystal 4The World Aquaculture Societyjohnc@was.orgfalseanrl65yqlzh3g1q0dme13067DD/MM/YYYY


Simon Chung*, Chen Su, Pin-Hsing Chou, Yu-Lun Liu, Yun-Long Tsai, Pei-Yu Alison Lee, and Hsiao-Fen Grace Chang
GenReach Biotechnology, Taichung City, 407, Taiwan

Polymerase chain reaction (PCR) for pathogen detection is a powerful tool to help maintain bio-security and improve overall shrimp production. The POCKITTM and POCKITTM PRO Nucleic Acid Analyzer (GeneReach) is a cost-effective and user-friendly tool to provide the same benefit at various aquaculture facilities. This system generates simple qualitative results from nucleic acid samples within one hour. Various iiPCR assays are available commercially for important shrimp pathogens, such as Enterocytozoon hepatopenaei (EHP), acute hepatopancreatic necrosis disease/early mortality syndrome, and white spot syndrome virus. EHP, a microsporidian parasite, has been associated with damages of hepatopancreas, malnutrition and growth retardation in shrimp, leading to significant economic losses. Here we evaluated the performance of the EHP iiPCR assay on POCKITTM method which could be used to minimize the potential threat by screening for the presence of EHP in water and shrimp.

The detection limit of the lyophilized EHV iiPCR assay was about 12 genome equivalents by testing serial dilutions of a standard plasmid DNA (Table 1). The assay did not react with Nosema ceranae (also a microsporidian), Vibrio parahaemolyticus, Vibrio harveyi, necrotizing hepatopancreatitis bacterium, WSSV and IHHNV. Performance of the iiPCR/POCKITTM system to detect EHP in shrimp was compared to a real-time PCR with 58 hepatopancreas and PL samples from Litopenaeus vannamei. Nucleic acids isolated by the taco Automatic Nucleic Acid Extraction System were tested side by side by the two PCR methods. Excellent agreement (96.55%; κ = 0.92) was found between the two assays.

Taken together, the established assay is useful tools for timely biosecurity and managing the EHP and other potential threats.

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