TY - JOUR
T1 - Amplification of tlh gene in other Vibrionaceae specie by specie-specific multiplex PCR of Vibrio parahaemolyticus
AU - Yáñez, Romina
AU - Bastías, Roberto
AU - Higuera, Gastón
AU - Salgado, Oscar
AU - Katharios, Pantelis
AU - Romero, Jaime
AU - Espejo, Romilio
AU - García, Katherine
N1 - Funding Information:
This research was supported by Fondecyt 11140257 , Fondecyt 11140412 , and Scientific Information Program/Fund for Scientific Journals Publishing , Year 2014, ID FP140010 .
Publisher Copyright:
© 2015 Pontificia Universidad Católica de Valparaíso. Production and hosting by Elsevier B.V. All rights reserved.
PY - 2015/11/15
Y1 - 2015/11/15
N2 - Background: The surveillance of Vibrio parahaemolyticus in the Chilean coast has been mainly performed by multiplex PCR amplification of three different hemolysin genes, which are specie-specific virulence factors. These genes are also employed in the determination of V. parahaemolyticus pathogenic load in seafood and for characterization of pathogenic strains associated to diarrhea cases in human. During environmental surveillance that we performed every summer, we occasionally observed a thermolabile hemolysin (tlh) PCR product of a slightly smaller size than expected, which was coincident with low loads of V. parahaemolyticus in the environment. In order to understand this observation, we probed the specificity of tlh primers for detection of V. parahaemolyticus at different bacterial loads and DNA concentrations. Results: Primers used for detection of V. parahaemolyticus specific tlh amplified a slightly smaller tlh gene, which is found in Vibrio alginolyticus and other related strains. These amplicons were observed when V. parahaemolyticus was absent or in undetectable loads in the environment. Conclusions: Surveillance of V. parahaemolyticus using tlh primers can be imprecise because amplification of a V. parahaemolyticus specific marker in V. alginolyticus and other related strains occurs. This situation complicates potentially the estimation of bacterial load in seafood, because do not ensure the correct identification of V. parahaemolyticus when his load is low. Additionally, it could complicate the tracking of outbreaks of V. parahaemolyticus infections, considering the genetic markers used would not be specie-specific.
AB - Background: The surveillance of Vibrio parahaemolyticus in the Chilean coast has been mainly performed by multiplex PCR amplification of three different hemolysin genes, which are specie-specific virulence factors. These genes are also employed in the determination of V. parahaemolyticus pathogenic load in seafood and for characterization of pathogenic strains associated to diarrhea cases in human. During environmental surveillance that we performed every summer, we occasionally observed a thermolabile hemolysin (tlh) PCR product of a slightly smaller size than expected, which was coincident with low loads of V. parahaemolyticus in the environment. In order to understand this observation, we probed the specificity of tlh primers for detection of V. parahaemolyticus at different bacterial loads and DNA concentrations. Results: Primers used for detection of V. parahaemolyticus specific tlh amplified a slightly smaller tlh gene, which is found in Vibrio alginolyticus and other related strains. These amplicons were observed when V. parahaemolyticus was absent or in undetectable loads in the environment. Conclusions: Surveillance of V. parahaemolyticus using tlh primers can be imprecise because amplification of a V. parahaemolyticus specific marker in V. alginolyticus and other related strains occurs. This situation complicates potentially the estimation of bacterial load in seafood, because do not ensure the correct identification of V. parahaemolyticus when his load is low. Additionally, it could complicate the tracking of outbreaks of V. parahaemolyticus infections, considering the genetic markers used would not be specie-specific.
KW - Hemolysin
KW - Multiplex PCR
KW - Pathogen surveillance
KW - Vibrio parahaemolyticus
KW - Virulence factor
UR - http://www.scopus.com/inward/record.url?scp=84946922086&partnerID=8YFLogxK
U2 - 10.1016/j.ejbt.2015.09.007
DO - 10.1016/j.ejbt.2015.09.007
M3 - Article
AN - SCOPUS:84946922086
VL - 18
SP - 459
EP - 463
JO - Electronic Journal of Biotechnology
JF - Electronic Journal of Biotechnology
SN - 0717-3458
IS - 6
ER -