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IQ2000 AHPND/EMS Toxin 1 Detection and Prevention System

 

Introduction

Acute hepatopancreatic necrosis disease (AHPND), also known as early mortality syndrome (EMS), is an important emerging shrimp disease. Affected shrimp would stop feeding, appear lethargic, and eventually die. Causing large-scale shrimp die-offs, AHPND/EMS has led to huge economic losses to shrimp farming industry.

Typical signs of AHPND/EMS infection, found mainly on the digestive tracts of shrimp, include empty gastrointestinal (GI) tract and abnormal shrunken whitish hepatopancreases. Damage in the hepatopancreatic system, including sloughing of hepatopancreas tubule epithelial cells, results in weakness and death in shrimp. The involvement of Vibrio parahaemolyticus (V. parahaemolyticus) containing a virulence-associated plasmid in shrimp or farming environment has been reported. The AHPND plasmid carries the virulence factor, toxin 1, which could damage the shrimp’s hepatopancreases. Toxin 1 is a complex of two proteins, namely PirA and PirB.

Technology transferred from the University of Arizona, Prof. Donald Lightner’s group and National Cheng Kung University, Prof. Chu-Fang Lo’s group, GeneReach has successfully developed the IQ2000™ AHPND/EMS Toxin 1 Detection and Prevention System to detect the virulence factor, toxin 1, and an AHPND/EMS-associated plasmid marker (the most stable region of the AHPND plasmid). This IQ2000™ Systems serves as a tool to detect and differentiate between the AHPND plasmids with and without the toxin 1 virulence factor to facilitate the implementation of shrimp disease control program in the shrimp farming industry.

With a great deal of research ongoing on AHPND/EMS, GeneReach reserves the right to modify and update the IQ2000™ AHPND/EMS Toxin 1 Detection & Prevention System accordingly.

Specifications:

  • Packing size: 200 reactions.
  • Detection Limit: 100 copies/reaction.
  • Quantitative capability: 3 different levels of infection can be differentiated.
  • Sample throughput: for 40 samples, from sample preparation to final results require 2.5 to 3 hours.
  • Built-in internal control system: eliminate false negative results from failed extraction or reaction.
  • Quantified positive standard: monitor the sensitivity of detection.

An example of the results is shown and explained below:

Lane 1: P(+) control, 2,000 copies/reaction
Lane 2: P(+) control, 200 copies/reaction
Lane 3: NTC(yeast tRNA or ddH2O)
Lane 4: Sample with toxin 1, virulent AHPND plasmid
Lane 5: Sample with non-virulent AHPND plasmid with toxin 1 deletion
Lane 6: Sample with PCR inhibitor
Lane 7: AHPND plasmid negative sample
Lane M: Molecular weight markers, 848bp, 630bp, and 333bp

Applications:

  • This reagent is intended for testing fresh, frozen, and ethanol-preserved samples, e.g. post larvae (PL), stomach, Mid-gut, hepatopancrease, and other samples such as pond water and feed organism (artemia, oyster, squid and blood worm).
  • Ideal for the specific pathogen-free (SPF) animal selection, evaluation of culture environment, grow-out monitoring, and quarantine service.

 

 
 
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