Quantification of the bacterial agent of necrotizing hepatopancreatitis (NHP-B) by real-time PCR and comparison of survival and NHP load of two shrimp populations

Luis Fernando Aranguren, Kathy F.J. Tang, Donald V. Lightner

Research output: Contribution to journalArticle

13 Scopus citations

Abstract

A real-time quantitative PCR (qPCR) assay was developed using a TaqMan probe to detect and quantify the necrotizing hepatopancreatitis bacterium (NHP-B) in Penaeus vannamei. A pair of primers which amplify a 67bp DNA fragment and a TaqMan probe were selected from the 16S rRNA gene of NHP-B genome. A positive control plasmid DNA was used to demonstrate that the NHP qPCR assay has a detection limit of 100 copies and a log-linear range up to 108 copies. An NHP-B challenge test using two different populations, Colombia (COL) and Specific Pathogen Free KONA line (KONA), was conducted. Higher final survival and a better survival curve were found in the COL population. No differences in NHP-B bacterial load were found in hepatopancreas (HP) analyzed from the survivors from each population. Sensitivity of the qPCR test was much higher than the conventional PCR (100 copies vs 1×105 copies).

Original languageEnglish (US)
Pages (from-to)187-192
Number of pages6
JournalAquaculture
Volume307
Issue number3-4
DOIs
StatePublished - Sep 1 2010

Keywords

  • Genetic selection
  • Necrotizing hepatopancreatitis
  • Penaeus vannamei
  • Real-time PCR

ASJC Scopus subject areas

  • Aquatic Science

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