Detection of Vibrio parahaemolyticus in shellfish by use of multiplexed real-time PCR with TaqMan fluorescent probes

Applied and Environmental Microbiology
Linda N Ward, Asim K Bej

Abstract

We developed a multiplexed real-time PCR assay using four sets of gene-specific oligonucleotide primers and four TaqMan probes labeled with four different fluorophores in a single reaction for detection of total and pathogenic Vibrio parahaemolyticus, including the pandemic O3:K6 serotype in oysters. V. parahaemolyticus has been associated with outbreaks of food-borne gastroenteritis caused by the consumption of raw or undercooked seafood and therefore is a concern to the seafood industry and consumers. We selected specific primers and probes targeting the thermostable direct hemolysin gene (tdh) and tdh-related hemolysin gene (trh) that have been reported to be associated with pathogenesis in this organism. In addition, we targeted open reading frame 8 of phage f237 (ORF8), which is associated with a newly emerged virulent pandemic serotype of V. parahameolyticus O3:K6. Total V. parahaemolyticus was targeted using the thermolabile hemolysin gene (tlh). The sensitivity of the combined four-locus multiplexed TaqMan PCR was found to be 200 pg of purified genomic DNA and 10(4) CFU per ml for pure cultures. Detection of an initial inoculum of 1 CFU V. parahaemolyticus per g of oyster tissue homogenate was possible after overnight e...Continue Reading

References

May 1, 1977·Applied and Environmental Microbiology·J E HobbieS Jasper
Sep 1, 1987·Kansenshōgaku zasshi. The Journal of the Japanese Association for Infectious Diseases·S HondaT Miwatani
Jan 1, 1982·Pharmacology & Therapeutics·Y Takeda
Jan 1, 1980·Annual Review of Microbiology·P A BlakeD G Hollis
Oct 1, 1994·Clinical Microbiology Reviews·S R Rippey
Jul 15, 1998·Current Microbiology·C W BrasherA K Bej
Feb 6, 1999·MMWR. Morbidity and Mortality Weekly Report·UNKNOWN Centers for Disease Control and Prevention (CDC)
Jun 2, 2000·Journal of Clinical Microbiology·H NasuT Honda
Sep 23, 2000·JAMA : the Journal of the American Medical Association·N A DanielsF J Angulo
Jun 1, 2001·Emerging Infectious Diseases·T IidaT Honda
Feb 2, 2002·Systematic and Applied Microbiology·S ChaiyananR R Colwell
Feb 16, 2002·Methods : a Companion to Methods in Enzymology·C T WittwerK S Elenitoba-Johnson
Sep 12, 2002·Journal of Microbiological Methods·V R LungeC A Batt
Oct 2, 2002·Clinical Infectious Diseases : an Official Publication of the Infectious Diseases Society of America·Israel PotasmanMajed Odeh
Mar 7, 2003·Applied and Environmental Microbiology·Angelo DePaolaDavid W Cook
Mar 26, 2003·Journal of Microbiological Methods·George M BlackstoneAngelo DePaola
Oct 9, 2003·Journal of Clinical Microbiology·Masatoshi OkuraHaruo Watanabe
Oct 14, 2003·FEMS Microbiology Letters·Antonio Lozano-LeónJaime Martínez-Urtaza
Nov 8, 2003·Journal of Clinical Microbiology·Hiroshi FukushimaRyotaro Seki
Jan 9, 2004·Applied and Environmental Microbiology·Gitika PanickerAsim K Bej
Feb 11, 2004·Kansenshōgaku zasshi. The Journal of the Japanese Association for Infectious Diseases·Mitsugu YamazakiYutaka Miyazaki
Oct 6, 2005·Applied and Environmental Microbiology·Gitika Panicker, Asim K Bej

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Citations

Jun 5, 2013·Archives of Virology·Martín Ramírez-OrozcoSergio F Martínez-Díaz
Dec 13, 2012·World Journal of Microbiology & Biotechnology·Piyanuch PrompamornParin Chaivisuthangkura
Jul 9, 2008·Food Additives & Contaminants. Part A, Chemistry, Analysis, Control, Exposure & Risk Assessment·A Obrepalska-SteplowskaJ Nawrot
Jan 16, 2007·Clinical Microbiology Reviews·G Balakrish NairDavid A Sack
Jan 1, 2008·Microbes and Environments·Youhei Fukui, Tomoo Sawabe
Dec 3, 2014·Foodborne Pathogens and Disease·Honghu SunQinghua Hu
Jun 22, 2011·Natural Computing·Fusheng Xiong, Wayne D Frasch
Feb 10, 2016·Frontiers in Microbiology·Ruichao LiSheng Chen
Oct 18, 2012·International Journal of Food Microbiology·Min ChenLu Feng
Oct 30, 2015·Environmental Science and Pollution Research International·Chang-Ho KangJae-Seong So
Jan 10, 2009·International Journal of Food Microbiology·V Blanco-AbadJ Martinez-Urtaza
Dec 25, 2008·International Journal of Food Microbiology·Anuj TyagiIndrani Karunasagar
Apr 29, 2008·Journal of Clinical Virology : the Official Publication of the Pan American Society for Clinical Virology·Manfred WeidmannFrank T Hufert
May 20, 2015·Journal of the Science of Food and Agriculture·Xi DengZhenqiang Chen
Mar 24, 2015·Frontiers in Microbiology·Rongzhi WangShihua Wang
Oct 23, 2016·Infection, Genetics and Evolution : Journal of Molecular Epidemiology and Evolutionary Genetics in Infectious Diseases·Xin LuBiao Kan
Sep 17, 2010·International Journal of Food Microbiology·A Robert-PillotM L Quilici
Jun 5, 2018·Canadian Journal of Microbiology·Deshun XuLiping Chen
Jun 15, 2011·Acta Veterinaria Hungarica·Urška HenigmanDarja Barlič-Maganja
Dec 31, 2019·The Canadian Journal of Infectious Diseases & Medical Microbiology = Journal Canadien Des Maladies Infectieuses Et De La Microbiologie Médicale·Bing WuHongling Wang
May 12, 2020·World Journal of Microbiology & Biotechnology·Karanth Padyana AnupamaBiswajit Maiti

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