Assay performance during validation of freezing channel catfish Ictalurus punctatus (Rafinesque) infected with a Gram‐negative bacterium |
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| Authors: | Julie Bebak Craig Shoemaker Covadonga Arias Phillip Klesius |
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| Institution: | 1. USDA ARS AAHRU, Auburn, AL, USA;2. Department of Fisheries and Allied Aquaculture, Auburn University, Auburn, AL, USA |
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| Abstract: | Recovery of bacteria from infected fish during population sampling can be affected by factors including the type of assay, method of specimen preservation and concentration of bacteria present. Consequently, before use in field sampling, methods should be validated. The three objectives of this study were, first, to determine whether a channel catfish Ictalurus punctatus (Rafinesque) fingerling classified as positive for Gram‐negative Edwardsiella ictaluri infection according to bacterial culture before freezing was also classified as positive after freezing, second, to determine how direct culture from the kidney (DIRECT), culture of homogenate (HOMOG) and standard PCR (PCR) agree with bacterial culture in terms of classifying fish as positive or negative and third, to estimate diagnostic sensitivity (dSe) and diagnostic specificity (dSp) for DIRECT, HOMOG and PCR. In fresh and frozen fish, as bacterial concentration decreased, the ability of each assay to detect positive fish also decreased, especially when there were <104 colony‐forming units per gram (CFU g?1) tissue. HOMOG proved to be the most reliable at correctly classifying catfish, whether they were subclinically or clinically infected. PCR assay was the least reliable. Overall, values for this study population for dSe were 0.66, 0.92 and 0.43, and for dSp were 0.86, 0.91 and 0.95, for DIRECT, HOMOG and PCR respectively. |
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| Keywords: | validation freezing Edwardsiella catfish PCR bacterial culture |
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