A Real-Time Polymerase Chain Reaction Assay of the BacteriumEdwardsiella ictaluriin Channel Catfish
- 1 March 2003
- journal article
- Published by Oxford University Press (OUP) in Journal of Aquatic Animal Health
- Vol. 15 (1) , 80-86
- https://doi.org/10.1577/1548-8667(2003)015<0080:arpcra>2.0.co;2
Abstract
A rapid (4.5-h) and sensitive assay based on polymerase chain reaction (PCR) was developed to facilitate the early detection of Edwardsiella ictaluri in channel catfish Ictalurus punctatus. A 129-base-pair fragment of a sequence specific to E. ictaluri was amplified with both standard and real-time (quantitative) PCR. The sensitivity of detection was determined to be as low as the equivalent of 2.5 cells in DNA samples from both E. ictaluri cells and mixtures of blood from noninfected catfish and E. ictaluri cells. Infection levels (as determined by real-time PCR) in blood from experimentally challenged fish were compared with brain–heart-infusion-cultured bacterial colony counts to assess the accuracy of the PCR assay. The PCR-based detection level (the equivalent of 105–108 cells/mL) was comparable to that of traditional culturing techniques (106–107 cells/mL). In future applications, this assay will be applied in a comprehensive breeding program to select channel catfish that are resistant to ...Keywords
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