{"refrec":{"BRefID":246903,"RR":"<b>Taminiau, B.; Korsak, N.; Lemaire, C.; Delcenserie, V.; Daube, G.</b> (2014). Validation of real-time PCR for detection of six major pathogens in seafood products. <i>Food Control 44</i>: 130-137. <a href=\"http://dx.doi.org/10.1016/j.foodcont.2014.03.031\" target=\"_blank\">dx.doi.org/10.1016/j.foodcont.2014.03.031</a>","BEntID":238603,"PublicFlag":1,"CheckedFlag":1,"wosflag":1,"vabbflag":1,"RefStringPartII":". <i>Food Control 44</i>: 130-137. <a href=\"https://dx.doi.org/10.1016/j.foodcont.2014.03.031\" target=\"_blank\">https://dx.doi.org/10.1016/j.foodcont.2014.03.031</a>","DocTypID":8,"DocType":"Journal article","MarineFlag":1,"FreshFlag":0,"BrackishFlag":0,"TerrestrialFlag":0,"Authorstring":"Taminiau, B.; Korsak, N.; Lemaire, C.; Delcenserie, V.; Daube, G.","OrigTitleTranslFlag":0,"Authorstringtrunc":"Taminiau, B. <i>et al.</i>","Englishabstract":"Seafood can pose a public health concern to consumers. It is often consumed raw and may be contaminated with several foodborne pathogens. In order to guarantee the safety of seafood, real-time polymerase chain reaction (PCR) protocols may be used as these enable results to be provided within 24 h. The first goal of our work was to develop real-time PCR protocols enabling the detection of six foodborne pathogens that may be present in seafood products (<i>Campylobacter jejuni</i>, <i>Campylobacter coli</i>, enterohemorrhagic <i>Escherichia coli</i>, <i>Salmonella</i> spp., <i>Vibrio parahaemolyticus</i>, and <i>Vibrio vulnificus</i>). The corresponding gene targets were: <i>50S/VS1</i>, <i>rfbE</i>, <i>ttr</i>, <i>tlh</i>, and <i>vvp</i>. A multiplex PCR was also developed to detect the virulence genes of <i>V. parahaemolyticus</i>: <i>tdh</i> and <i>trh</i>. A total of 420 bacterial strains belonging to four different genera/strains were used in this study. Sensitivity and specificity were always 100%, except in the case of <i>Salmonella</i> spp., where three strains were not detected by our PCR protocols. The second objective of our work was to assess the detection limit of our real-time PCR protocols on artificially contaminated seafood products (raw shrimps, cooked shrimps, and raw mussels), purchased in public stores. Six different levels of contamination were assayed in four replicates for each matrix. The real-time PCR protocols enabled a better level of detection than the ISO methods, except for <i>Salmonella</i> in raw shrimps and for <i>V. vulnificus</i> in shrimps (raw and cooked). The estimated level of detection was between 1 and 47 cfu/25 g sample for the ISO norms and between 1 and 315 cfu/25 g sample for the real-time PCR protocols tailored in our work. The real-time PCRs developed in our work allowed for good selectivity, sensitivity, and specificity. The sensitivity on seafood products was estimated at a level of 100%, except for <i>Salmonella</i> (97%). In the spiking assays, the levels of detection were lower with the real-time PCR protocol than those obtained with the ISO method. This was not the case for <i>V. vulnificus</i> in raw and cooked shrimps and for <i>Salmonella</i> in raw shrimps. These real-time PCR protocols appear to be good alternative methods for surveillance of seafood products to ensure the absence of foodborne pathogens. One additional conclusion is that laboratories have to use enrichment media that are compatible with those recommended by ISO standards. This may facilitate the isolation of the pathogen if the real-time PCR protocol gives a suspect positive signal during the first step of the seafood analysis.","AbstractOtherLang":null,"BibLvlCode":"AS","StandardTitle":"Validation of real-time PCR for detection of six major pathogens in seafood products","OrigTitleLangCode":"en","OrigTitleLangCodeExtended":"eng","OrigTitleLangID":15,"DateLastModified":{"date":"2024-12-10 01:33:17.368041","timezone_type":1,"timezone":"+01:00"},"UserAccessRight":null,"UserAccID":null,"AuthorKeywords":"Seafood; Real-time PCR; Campylobacter; EHEC; Salmonella; Vibrios","OtherDescriptors":null,"Notes":null,"AnaPub":2014,"MonPub":null,"DateUpdate":"2015-10-28","DateCreate":"2015-05-21","SecASFANote":null,"ConfID":null,"PeerRev":1,"VlizCoreFlag":1,"WoScode":"WOS:000338619900017","VABBcode":null,"OpenAcc":0,"DOI":"10.1016/j.foodcont.2014.03.031"},"refs":null,"anarec":{"AnaID":246903,"PubliDate":2014,"Pagination":"130-137","XtraPublOfAnaID":null,"ISBN":null,"Volume":"44","Issue":null,"BRefMon":null,"BRefMonRR":null,"BRefXtra":null,"BRefXtraRR":null,"SerBRefID":220415,"SerRR":"Food Control. 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