2004
DOI: 10.3354/dao059195
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Quantitative real-time RT-PCR demonstrates that handling stress can lead to rapid increases of gill-associated virus (GAV) infection levels in Penaeus monodon

Abstract: Gill-associated virus (GAV) of the black tiger prawn Penaeus monodon has been implicated as a cause of periodic production losses in Australia since 1996. We report here the development of a real-time quantitative RT-PCR (qRT-PCR) for GAV. A dilution series of in vitro transcribed RNA was used to determine the sensitivity limit of the qRT-PCR and as a standard for GAV quantification. A linear relationship between cycle threshold (Ct) values and input RNA was obtained over a wide concentration range between 4.8… Show more

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Cited by 63 publications
(29 citation statements)
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“…GAV loads of Novacq TM fed shrimp were on average lower in individual shrimp over the duration of the experimental bioassay when they received the stress of a saline in injection only. It has been previously demonstrated that handling stress such as that caused by inserting a needle into P. monodon can result in heightened GAV loads of unselected lines (de la Vega et al, 2004). Although less pronounced, the same inferred 'fitness' measured as generally lower GAV loads in individuals throughout the duration of the experimental bioassay, appears amongst Novacq TM fed shrimp that received both the handling stress of injection and the GAV challenge.…”
Section: Gav Load Comparisonmentioning
confidence: 91%
“…GAV loads of Novacq TM fed shrimp were on average lower in individual shrimp over the duration of the experimental bioassay when they received the stress of a saline in injection only. It has been previously demonstrated that handling stress such as that caused by inserting a needle into P. monodon can result in heightened GAV loads of unselected lines (de la Vega et al, 2004). Although less pronounced, the same inferred 'fitness' measured as generally lower GAV loads in individuals throughout the duration of the experimental bioassay, appears amongst Novacq TM fed shrimp that received both the handling stress of injection and the GAV challenge.…”
Section: Gav Load Comparisonmentioning
confidence: 91%
“…The nested PCR did not extend the detection limit beyond 2 to 6 RNA copies but amplicon yields were significantly greater and clearly detected in agarose gels. The RT-nested PCR sensitivity thus approaches the theoretical limit of 1 RNA copy and compares favourably to a similar test to detect GAV (Cowley et al 2000a, de la Vega et al 2004) and realtime RT-PCR tests for GAV (de la Vega et al 2004) and other RNA viruses including YHV and TSV (Dhar et al 2002). Moreover, as very few (less than 0.2%) nucleotide changes occurred in MoV RT-PCR products amplified from P. monodon from eastern Australia, the test should be quite robust in detecting minor quasispecies variants present in this prawn population.…”
Section: Discussionmentioning
confidence: 99%
“…In wild-captured P. monodon broodstock, repeated handling stress associated with bleeding has also been noted to markedly increase GAV infection levels in haemocytes [155].…”
Section: Mechanisms Of Protectionmentioning
confidence: 99%