scholarly journals Molecular Detection Methods for Vibrio parahaemolyticus in Seafood

2015 ◽  
Vol 89 (4) ◽  
pp. 445-451 ◽  
Author(s):  
Tomohiro NISHIO ◽  
Kayoko OHTSUKA ◽  
Midori ODA ◽  
Kanji SUGIYAMA ◽  
Yukiko HARA-KUDO
2012 ◽  
Vol 30 (1) ◽  
pp. 105-111 ◽  
Author(s):  
Jessica L. Jones ◽  
Yukiko Hara-Kudo ◽  
Jeffrey A. Krantz ◽  
Ronald A. Benner ◽  
Amy B. Smith ◽  
...  

2018 ◽  
Vol 15 (1) ◽  
Author(s):  
Ameh S. James ◽  
Shawn Todd ◽  
Nina M. Pollak ◽  
Glenn A. Marsh ◽  
Joanne Macdonald

2006 ◽  
Vol 72 (3) ◽  
pp. 2031-2042 ◽  
Author(s):  
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 104 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 enrichment, which resulted in a concentration of 3.3 × 109 CFU per ml. Use of this method with natural oysters resulted in 17/33 samples that were positive for tlh and 4/33 samples that were positive for tdh. This assay specifically and sensitively detected total and pathogenic V. parahaemolyticus and is expected to provide a rapid and reliable alternative to conventional detection methods by reducing the analysis time and obviating the need for multiple assays.


2009 ◽  
Vol 24 (4) ◽  
pp. 215-222 ◽  
Author(s):  
Apostolos Zaravinos ◽  
Ioannis N. Mammas ◽  
George Sourvinos ◽  
Demetrios A. Spandidos

2007 ◽  
Vol 56 (4) ◽  
pp. 487-490 ◽  
Author(s):  
Iona M. C. Martin ◽  
Ellie Foreman ◽  
Vicky Hall ◽  
Anne Nesbitt ◽  
Greta Forster ◽  
...  

Isolation of Neisseria gonorrhoeae is currently the gold standard for the definitive diagnosis of gonorrhoea and for use in medico-legal cases in the UK. Molecular detection methods are used increasingly but are untested as evidence of infection in a court of law. An isolate of N. gonorrhoeae was obtained from a child and an article of clothing from an adult male who was suspected of sexual abuse of the child. Biochemical and immunological tests were used to confirm the isolate as N. gonorrhoeae. Amplification by PCR using two targets, cppB and ompIII, was used both as further confirmation of the isolate and to detect the presence of gonococcal-specific DNA from the clothing. The relationship of the gonococcal DNA from the child and the adult was investigated using genotyping (N. gonorrhoeae multi-antigen sequence typing; NG-MAST), including a nested PCR for the por gene. Both samples were indistinguishable by NG-MAST and shared the same sequence type, 403. This is the first report of molecular detection and genotyping of N. gonorrhoeae on an article of clothing, which resulted in conviction of the man for sexual assault.


2013 ◽  
Vol 112 (5) ◽  
pp. 2087-2093 ◽  
Author(s):  
Zhibing Lin ◽  
Jie Cao ◽  
Houshuang Zhang ◽  
Yongzhi Zhou ◽  
Mingjun Deng ◽  
...  

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