The MaxSignal® Shrimp WSSV Detection Kit uses an end-point Polymerase Chain Reaction (PCR) assay to screen for WSSV in fresh or frozen shrimp or other potentially infectious material. White Spot Syndrome Virus (WSSV) is claimed to be the most serious viral pathogen that affects cultivated shrimp worldwide. The MaxSignal® Shrimp WSSV Detection Kit contains 2 pairs of oligonucleotide primers targeting nested regions of the WSSV VP466 gene (accession # EF564812), which encodes a structural protein located in the nucleocapsid. The WSSV amplicon produced using the WSSV-483 primers in the kit is 483 bp and the amplicon produced using the WSSV-388 primers is 388 bp. These WSSV amplicons have no significant homology to any other viral sequence. The detection limit for the synthetic WSSV template included in the kit is approximately 50 copies using the standard 1-step PCR protocol, and approximately 5 copies using the 2-step nested PCR protocol. For detection of WSSV in infected shrimp, positive signals have been obtained using the 1-step PCR protocol with input amounts of approximately 50 femtograms (50 x 10-15 gm) of total nucleic acid extracted from shrimp that were experimentally infected with high levels of the WSSV virus. The Shrimp Positive Control primers included in the kit were designed to the actin 1 gene (Accession # AF100986) from Penaeus monodon and will produce an amplicon of 249 bp from P monodon and also from Penaeus vannamei. The Shrimp Positive Control primers are used to demonstrate successful extraction of DNA that is able to support PCR. Input sample for this kit is total DNA or total nucleic acid extracted from solid tissue or derived from liquid tissues such as hemolymph; purification of viral particles for analysis is not required. To maximize user convenience, the reagents for PCR (buffer, dNTPs, and thermostable polymerase) are provided as a 5x MasterMix. The MasterMix also contains components to allow the PCR to be directly loaded for gel analysis, without adding gel-loading solution.
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