TY - JOUR
T1 - 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
AU - Aranguren, Luis Fernando
AU - Tang-Nelson, Feng-Jyu
AU - Lightner, Donald V
N1 - Funding Information:
This work was supported by the Gulf Coast Research Laboratory Consortium Marine Shrimp Farming Program, USDA Grant No. USMSFD 2009/19851-UAZ and the S-K grant No: NA09NMF4270102 , NOAA . We thank to Dr. Marcela Salazar and Dr. James Cock for the critical reviewing of this manuscript, Linda Nunan and Brenda Noble for providing technical assistance, Rita Redman for histological services and Dr. H. Harris for providing the NHP-B infected tissue.
PY - 2010/9
Y1 - 2010/9
N2 - 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).
AB - 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).
KW - Genetic selection
KW - Necrotizing hepatopancreatitis
KW - Penaeus vannamei
KW - Real-time PCR
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U2 - 10.1016/j.aquaculture.2010.07.022
DO - 10.1016/j.aquaculture.2010.07.022
M3 - Article
AN - SCOPUS:77956267841
SN - 0044-8486
VL - 307
SP - 187
EP - 192
JO - Aquaculture
JF - Aquaculture
IS - 3-4
ER -