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Detection of Channel Catfish Virus in Adult Channel Catfish by Use of a Nested Polymerase Chain Reaction
Author(s) -
Baek YoungSook,
Boyle John A.
Publication year - 1996
Publication title -
journal of aquatic animal health
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.507
H-Index - 52
eISSN - 1548-8667
pISSN - 0899-7659
DOI - 10.1577/1548-8667(1996)008<0097:doccvi>2.3.co;2
Subject(s) - catfish , ictalurus , ethidium bromide , biology , polymerase chain reaction , nested polymerase chain reaction , broodstock , microbiology and biotechnology , primer (cosmetics) , real time polymerase chain reaction , dna , virology , fishery , gene , genetics , fish <actinopterygii> , aquaculture , chemistry , organic chemistry
Optimal polymerase chain reaction (PCR) conditions were determined with nested primers targeted to the DNA polymerase gene of channel catfish virus (CCV). Serial dilutions of CCV DNA in the presence or absence of DNA of channel catfish Ictalurus punctatus were used as templates throughout the optimization procedure. The final PCR product was detected by visualization in acrylamide gels stained with ethidium bromide. Fewer than 10 copies of CCV DNA (about 1 fg) could be detected in the presence of 10 8 times as much channel catfish DNA. The optimized two‐step PCR assay allowed detection of latent CCV in blood samples from healthy broodfish. Our results make possible the development of a routine testing procedure for catfish broodstock so that CCV can be eliminated in young catfish.