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IQ2000 IHHNV (nested) Detection and Prevention System

 

Introduction

Infectious Hypodermal and Hematopoietic Necrosis Virus (IHHNV) has been wildly reported as one of the major viral diseases in shrimp culture industry. It is known to be transmitted both vertically and horizontally. This single strand, negative sense DNA virus can cause mass mortality for L. stylirostris and runt deformity syndrome (RDS) for L. vannamei. Recent research findings pointed out that there is a close correlation between IHHNV and muscle whitening syndrome. Although IHHNV may not affect the growth of P. monodon significantly, however, recent research results also pointed out that it will influence the survival rate of the monodon broodstocks. Thus, the economic loss cause by this virus has been underestimated for a long time.

Dr. Kathy F. J. Tang published the IHHNV-related sequence in the genome of P. monodom from Africa and Australia in late 2006. They developed a PCR method which can be used to differentiate the IHHNV sequence from the IHHNV-related one in 2007. Their findings told there is opportunity of false positive test results if tested by some commercial PCR test kit. After our tests on the P. monodon come from Africa and Australia by our IQ2000 IHHNV one step PCR kit, 5% false positive results were found. However, no false positive result is found on L. vannamei.

To ensure the accuracy of the test result on P. monodon especially from Africa and Australia, we developed the IQ2000 IHHNV (nested) Detection and Prevention System to fulfill the requirement of the customers who need more accurate and higher bio-security level.

With the new system, not only the false-positive results can be avoided, but also the detection sensitivity will be increased to 10 copies per reaction same as other IQ2000 nested PCR test kits. In addition, IQ2000 IHHNV is specific to detect Type 1 and Type 2 infectious IHHNV, and will not react with Type 3A and 3B IHHNV, which are non-infectious and have been found inserted into the genome of P. monodon from East Africa, Australia, and the western Indo-Pacific region.

Specifications:

  • Packing size: 200 reactions.
  • Detection Limit: 10 copies/reaction.
  • Quantitative capability: 3 different levels of infection can be differentiated.
  • Sample throughput: for 40 samples, from sample preparation to final results require 3.5 to 4 hours.
  • Built-in internal control system: eliminate false negative results from failed extraction or reaction.
  • Quantified positive standard: monitor the sensitivity of detection.
  • Adaptable to the Uni-IQ RT-PCR profile, suitable for multi-viral diagnosis.

An example of the results is shown and explained below:

Lane M:  Molecular weight markers, 848 bp, 630 bp, 333 bp
Lane 1:  IHHNV P(+) standard, 2,000 copies/reaction
Lane 2:  IHHNV P(+) standard, 200 copies/reaction
Lane 3:  IHHNV P(+) standard, 20 copies/reaction
Lane 4:  Sample of moderate IHHNV infection
Lane 5:  Sample of light IHHNV infection
Lane 6:  Sample of very light IHHNV infection
Lane 7:  IHHNV negative sample
Lane 8:  Negative control (Yeast tRNA or ddH2O)

Applications:

  • This reagent is intended for testing fresh, frozen, and ethanol-preserved samples, e.g. pleopod, and PL.
  • Ideal for the specific pathogen-free (SPF) animal selection, evaluation of culture environment, grow-out monitoring, and quarantine service.
 
 
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