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ARS Home » Northeast Area » Wyndmoor, Pennsylvania » Eastern Regional Research Center » Residue Chemistry and Predictive Microbiology Research » Research » Publications at this Location » Publication #316210

Research Project: DEVELOPMENT OF PREDICTIVE MICROBIAL MODELS FOR FOOD SAFETY AND THEIR ASSOCIATED USE IN INTERNATIONAL MICROBIAL DATABASES

Location: Residue Chemistry and Predictive Microbiology Research

Title: Evaluation of reference genes in Vibrio parahaemolyticus for gene expression analysis using quantitative RT-PCR

Author
item Ma, Yue-jiao - Shanghai University
item Xu, Xiao-yan - Shanghai University
item Sun, Xiaohong - Shanghai University
item Zhao, Yong - Shanghai University
item Pan, Ying-jie - Shanghai University
item Hwang, Cheng-an - Andy
item Wu, Vivian - University Of Maine

Submitted to: PLoS One
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 11/23/2015
Publication Date: 12/11/2015
Citation: Ma, Y., Xu, X., Sun, X., Zhao, Y., Pan, Y., Hwang, C., Wu, V. 2015. Evaluation of reference genes in Vibrio parahaemolyticus for gene expression analysis using quantitative RT-PCR. PLoS One. 10(12):e0144362. doi:10.1371/journal.pone.0144362.

Interpretive Summary: Vibrio parahaemolyticus is frequently implicated in foodborne illnesses linked to the consumption of contaminated seafood. This study evaluated the stability of 6 reference genes (16S rRNA, recA, rpoS, pvsA, pvuA, and gapdh) in 5 V. parahaemolyticus strains cultured at 15, 25, 37 and 42 degrees Celsius to identify a stable gene as a reference gene for qRT-PCR analysis. The results indicated that recA was the most stably expressed gene and it can be used as a reference gene for gene expression studies in V. parahaemolyticus.

Technical Abstract: Vibrio parahaemolyticus is a significant human pathogen capable of causing foodborne gastroenteritis associated with the consumption of contaminated raw or undercooked seafood. Quantitative RT-PCR (qRT-PCR) is a useful tool for studying gene expression in V. parahaemolyticus to characterize the virulence factors and understand the interactions between the pathogenicity and environment conditions. However, there is not a stable gene in V. parahaemolyticus that has been identified for use as a reference gene for qRT-PCR. This study evaluated the stability of 6 reference genes (16S rRNA, recA, rpoS, pvsA, pvuA, and gapdh) in 5 V. parahaemolyticus strains (O3:K6-clinical strain-tdh+, ATCC33846-tdh+, ATCC33847-tdh+, ATCC17802-trh+, and F13-environment strain-tdh+) cultured at different temperatures (15, 25, 37 and 42 degrees Celsius). Stability values were calculated using geNorm, NormFinder, BestKeeper, and Delta CT algorithms. The results indicated that recA was the most stably expressed gene in the 5 V. parahaemolyticus strains cultured at different temperatures. This study suggests that recA can be used as a reference gene for gene expression studies in V. parahaemolyticus.