Quantitative Polymerase Chain Reaction Assay for Largemouth Bass Virus

The use of quantitative polymerase chain reaction (QPCR) to test for largemouth bass virus (LMBV) was evaluated during a challenge experiment in which largemouth bass Micropterus salmoides were immersed in the type strain of LMBV. The real‐time PCR and cell culture methods were both used to measure...

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Bibliographic Details
Published in:Journal of aquatic animal health 2007-12, Vol.19 (4), p.226-233
Main Authors: Getchell, R. G., Groocock, G. H., Schumacher, V. L., Grimmett, S. G., Wooster, G. A., Bowser, P. R.
Format: Article
Language:English
Subjects:
Animals
Bass - virology
Capsid Proteins - genetics
Cell Line
DNA Primers - chemistry
DNA Virus Infections - diagnosis
DNA Virus Infections - veterinary
DNA Virus Infections - virology
Fish Diseases - diagnosis
Fish Diseases - virology
Freshwater
Largemouth bass virus
Micropterus salmoides
Polymerase Chain Reaction - methods
Polymerase Chain Reaction - veterinary
Ranavirus - genetics
Ranavirus - isolation & purification
Reference Values
Reproducibility of Results
Sensitivity and Specificity
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Summary:The use of quantitative polymerase chain reaction (QPCR) to test for largemouth bass virus (LMBV) was evaluated during a challenge experiment in which largemouth bass Micropterus salmoides were immersed in the type strain of LMBV. The real‐time PCR and cell culture methods were both used to measure LMBV present in the inoculum. Additional samples tested by QPCR included gill, gonad, kidney, liver, mucus, spleen, and swim bladder. A plasmid clone containing a 248‐base pair (bp) fragment of the major capsid protein gene (MCP*) was serially diluted and used as a standard to quantify the number of LMBV DNA copies present in the samples tested. A 62‐bp fragment of DNA located in MCP* was amplified in the real‐time PCR assay. This work has demonstrated the value of the QPCR assay in LMBV surveys.
ISSN:0899-7659
1548-8667
DOI:10.1577/H07-009.1