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A molecular method to detect and identify the native species of southwestern Atlantic Crassostrea (Mollusca: Ostreidae) Rev. Bras. Zool.
Ludwig,Sandra; Patella,Raquel; Stoiev,Sérgio; Castilho-Westphal,Gisela; Girotto,Marcus V. F; Ostrensky,Antonio.
Among oysters, species of Crassostrea (Sacco, 1897) are the most attractive to aquaculture. In Brazil, the genus is represented by C. rhizophorae (Guilding, 1828) and C. brasiliana (Lamarck, 1819). Because the maturation and breeding technology is not well developed for these species, aquaculturists need a reliable method to decide the correct time to place spat collectors in the field, and to identify both species, which are morphologically similar. In this study a specific Multiplex PCR protocol was developed, using one pair of universal primers from 18S rDNA as a positive control and a pair of specific primers for each target species. The sensitivity and specificity of the protocol was evaluated. It detected C. rhizophorae DNA in low concentrations, and...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Multiplex PCR; Oysters; Plankton; Specific primers.
Ano: 2011 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1984-46702011000400002
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A novel multiplex PCR for the simultaneous detection of Salmonella enterica and Shigella species BJM
Radhika,M.; Saugata,Majumder; Murali,H.S.; Batra,H.V..
Salmonella enterica and Shigella species are commonly associated with food and water borne infections leading to gastrointestinal diseases. The present work was undertaken to develop a sensitive and reliable PCR based detection system for simultaneous detection of Salmonella enterica and Shigella at species level. For this the conserved regions of specific genes namely ipaH1, ipaH, wbgZ, wzy and invA were targeted for detection of Shigella genus, S. flexneri, S. sonnei, S. boydii and Salmonella enterica respectively along with an internal amplification control (IAC). The results showed that twenty Salmonella and eleven Shigella spp., were accurately identified by the assay without showing non-specificity against closely related other Enterobacteriaceae...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Salmonella; Shigella; Multiplex PCR; Spiking.
Ano: 2014 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1517-83822014000200041
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Amplification of tlh gene in other Vibrionaceae specie by specie-specific multiplex PCR of Vibrio parahaemolyticus Electron. J. Biotechnol.
Yáñez,Romina; Bastías,Roberto; Higuera,Gastón; Salgado,Oscar; Katharios,Pantelis; Romero,Jaime; Espejo,Romilio; García,Katherine.
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 the detection of V....
Tipo: Journal article Palavras-chave: Hemolysin; Multiplex PCR; Pathogen surveillance; Virulence factor; Vibrio parahaemolyticus.
Ano: 2015 URL: http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582015000600012
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Clostridium perfringens isolate typing by multiplex PCR J. Venom. Anim. Toxins incl. Trop. Dis.
Ahsani,MR; Mohammadabadi,MR; Shamsaddini,MB.
Clostridium perfringens is an important pathogen that provokes numerous different diseases. This bacterium is classified into five different types, each of which capable of causing a different disease. There are various methods for the bacterial identification, many are labor-intensive, time-consuming, expensive and also present low sensitivity and specificity. The aim of this research was to identify the different types of C. perfringens using PCR molecular method. In this study, 130 sheep-dung samples were randomly collected from areas around the city of Kerman, southeastern Iran. After processing and culturing of samples, the produced colonies were morphologically studied, gram stain test was also carried out and the genera of these bacteria were...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Clostridium perfringens; Multiplex PCR; Biochemical tests; Genetic typing.
Ano: 2010 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1678-91992010000400006
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Conventional versus molecular tests (Multiplex PCR and PCR mecA gene) for detection of methicillin resistant Staphylococcus aureus BJM
Ribas,Rosineide Marques; Gontijo-Filho,Paulo Pinto; Darini,Ana Lúcia da Costa.
In this study, for detection of methicillin-resistant S. aureus (MRSA), a mecA multiplex PCR-based amplification was compared with the 1 µg oxacillin disk diffusion test, detection of minimal inhibitory concentration (MIC), and screening in agar with 4% NaCl and 6 µg/mL oxacillin. Among 24 isolates obtained from blood, mecA gene was detected in only 16 (66.7%) isolates by multiplex PCR. The MIC test showed a range of resistance to oxacillin from 0.19 to 512 µg/mL, among these isolates. Data obtained by screening and dilution tests showed that sensitivity to methicillin was 80.0% and 72.8%, respectively, when compared with the presence of mecA gene (multiplex). All isolates, including the negatives, when revaluated for mecA gene by PCR were positive....
Tipo: Info:eu-repo/semantics/article Palavras-chave: Multiplex PCR; MRSA; Beta-lactamase; MecA gene.
Ano: 2003 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1517-83822003000500012
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Correlation between API 50 CH and multiplex polymerase chain reaction for the identification of vaginal lactobacilli in isolates BJM
Brolazo,Eliane Melo; Leite,Domingos Silva; Tiba,Monique Ribeiro; Villarroel,Marina; Marconi,Camila; Simoes,Jose Antonio.
Identification of Lactobacillus sp. strains by phenotypic methods may lead to doubtful results possibly interfering in the reliability of the epidemiological and probiotics studies. Therefore this study aimed to determine the best methodology for the identification of the large diversity of lactobacilli species found in the vagina by comparing two techniques, one based on their biochemical profile and other employing molecular biology. A carbohydrate fermentation test (API 50 CH) was compared with multiplex polymerase chain reaction (PCR) for the identification of species of vaginal lactobacilli from 135 healthy women. The kappa index was used to evaluate agreement between the methods. Using the molecular technique, L. crispatus (32.6%), L. jensenii (25%)...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Carbohydrate fermentation test; Multiplex PCR; Identification; Vaginal lactobacilli; Vaginal ecosystem.
Ano: 2011 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1517-83822011000100028
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Detection by multiplex PCR of Staphylococcus aureus , S. intermedius and S. hyicus in artificially contaminated milk Ciência Rural
Gandra,Eliezer Avila; Fernandez,Maria Aparecida; Silva,Jorge Adolfo; Silva,Wladimir Padilha da.
ABSTRACT: This research aimed to detect coagulase-positive Staphylococcus (CPS) directly in samples of artificially contaminated milk, using multiplex PCR (mPCR). Standard and isolated bacterial strains of S. aureus, S. epidermidis, S. hyicus, S. intermedius, Listeria monocytogenes and Escherichia coli species were used, evaluating the specificity and detection limit of mPCR, for artificially contaminated UHT milk. Primers specific for the nuc gene (NUC1-NUC2 were used for S. aureus, NUC3-NUC4 for S. hyicus and NUC5-NUC6 for S. intermedius). It was possible to detect the three target species by mPCR, directly from bovine whole milk, with adequate specificity and acceptable detention limit for identification of coagulase-positive Staphylococcus (CPS) in...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Staphylococcus aureus; Staphylococcus hyicus; Staphylococcus intermedius; Multiplex PCR; Nuc gene.
Ano: 2016 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0103-84782016000801418
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Detection of the mecA gene and identification of Staphylococcus directly from blood culture bottles by multiplex polymerase chain reaction BJID
Rocchetti,Taisa Trevizani; Martins,Katheryne Benini; Martins,Patricia Yoshida Faccioli; Oliveira,Rogério Antonio de; Mondelli,Alessandro Lia; Fortaleza,Carlos Magno Castelo Branco; Cunha,Maria de Lourdes Ribeiro de Souza da.
ABSTRACT Introduction: Staphylococcus spp. – both S. aureus, including methicillin-resistant strains (MRSA) and coagulase negative staphylococci (CoNS) – are relevant agents of healthcare-associated infections. Therefore, the rapid recognition of MRSA and methicillin-resistant CoNS from blood stream infections is critically important for patient management. It is worth noting that inappropriate empiric therapy has been associated with higher in-hospital mortality. Material and methods: In this study we evaluated a multiplex polymerase chain reaction (multiplex PCR) standardized to detect Staphylococcus spp., S. aureus, and mecA gene-encoded oxacillin resistance directly from blood culture bottles. A total of 371 blood cultures with Gram-positive...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Staphylococcus spp.; Staphylococcus aureus; MRSA; MecA gene; Blood cultures; Multiplex PCR.
Ano: 2018 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1413-86702018000200099
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Development of four EST-SSR multiplex PCRs in the Pacific oyster (Crassostrea gigas) and their validation in parentage assignment ArchiMer
Li, Ronghua; Li, Qi; Cornette, Florence; Degremont, Lionel; Lapegue, Sylvie.
We report four highly informative multiplex PCRs developed from twelve previously described ESTSSRs in Crassostrea gigas. We evaluated and validated these multiplex PCRs in twelve full-sib families. The average allelic richness and the polymorphism information content (PIC) were 11.1 and 0.811 respectively. The combined power of exclusion was greater than 99.99% using all four multiplex assays. A hundred and forty three tests of segregation ratios revealed 11 significant departures from expected Mendelian ratios. The frequency of null alleles was estimated as 4.9% of all the alleles segregating based on a within-family analysis of Mendelian segregation patterns. Parentage analysis of real offspring demonstrated that 97% of all offspring were unambiguously...
Tipo: Text Palavras-chave: Pacific oyster; Crassostrea gigas; Microsatellite; EST; Multiplex PCR; Parentage assignment.
Ano: 2010 URL: http://archimer.ifremer.fr/doc/00013/12419/9999.pdf
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Innovative molecular approach to the identification of Colossoma macropomum and its hybrids Anais da ABC (AABC)
Gomes,Fátima; Schneider,Horacio; Barros,Claudene; Sampaio,Dioniso; Hashimoto,Diogo; Porto-Foresti,Fábio; Sampaio,Iracilda.
Tambaqui (Colossoma macropomum) is the fish species most commonly raised in the Brazilian fish farms. The species is highly adaptable to captive conditions, and is both fast-growing and relatively fecund. In recent years, artificial breeding has produced hybrids with Characiform species, known as "Tambacu" and "Tambatinga". Identifying hybrids is a difficult process, given their morphological similarities with the parent species. This study presents an innovative molecular approach to the identification of hybrids based primarily on Multiplex PCR of a nuclear gene (α-Tropomyosin), which was tested on 93 specimens obtained from fish farms in northern Brazil. The sequencing of a 505-bp fragment of the Control Region (CR) permitted the identification of the...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Pisciculture; Multiplex PCR; Mitochondrial DNA; Genetic bottleneck.
Ano: 2012 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0001-37652012000200020
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Isolation and multiplex genotyping of polymorphic microsatellite DNA markers in the snakehead murrel, Channa striata Genet. Mol. Biol.
Jamsari,Amirul Firdaus Jamaluddin; Min-Pau,Tan; Siti-Azizah,Mohd Nor.
Seven polymorphic microsatellite loci were isolated and characterized for the snakehead murrel, Channa striata (Channidae), a valuable tropical freshwater fish species. Among 25 specimens collected from Kedah state in Malaysia, the number of alleles per locus ranged from 2 to 7. Observed and expected heterozygosities ranged from 0.120 to 0.880 and 0.117 to 0.698, respectively. A single locus (CS1-C07) was significantly deviated from Hardy-Weinberg equilibrium after Bonferroni correction. These novel markers would be useful for population genetic studies of the C. striata.
Tipo: Info:eu-repo/semantics/article Palavras-chave: Channa striata; Population genetics; Microsatellite; Multiplex PCR.
Ano: 2011 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1415-47572011000200029
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Methods for the evaluation of antibiotic resistance in Lactobacillus isolated from fermented sausages Ciência Rural
Wolupeck,Hanna Lethycia; Morete,Crisley Aparecida; DallaSanta,Osmar Roberto; Luciano,Fernando Bittencourt; Madeira,Humberto Maciel França; Macedo,Renata Ernlund Freitas de.
ABSTRACT: The present study aimed to assess the antibiotic resistance in 54 indigenous Lactobacillus plantarum isolated from artisanal fermented sausages. The confirmation of the strain species was performed by multiplex-PCR assay. Antibiotic resistance was assessed by disk diffusion (DD) and Minimum Inhibitory Concentration (MIC) methods. Of 54 L. plantarum, 44 strains were genotypically confirmed as L. plantarum and 3 as Lactobacillus pentosus. The highest resistance rates were to ampicillin and streptomycin. The highest susceptibility rates were shown to tetracycline, chloramphenicol and penicillin G. None of the strains showed multidrug resistance. Resistance rates by DD and MIC were not different (P>0.05) for ampicillin, chloramphenicol,...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Meat product; Multiplex PCR; Probiotic; Susceptibility.
Ano: 2017 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0103-84782017000800452
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Molecular characterization of Rhodococcus equi isolates of horse breeding farms from an endemic region in South of Brazil by multiplex PCR BJM
Krewer,Cristina da Costa; Spricigo,Dênis Augusto; Botton,Sônia de Avila; Costa,Mateus Matiuzzi da; Schrank,Irene; Vargas,Agueda Castagna de.
Rhodococcus equi is a gram-positive coco-bacillus and an intracellular opportunistic pathogen which causes pneumonia in foals. It is widely detected in environment and has been isolated from several sources, as soil, feces and gut from health and sick foals. The goal of this study was to characterize the epidemiological status (endemic, sporadic or no infection) of horse breeding farms from Bage County in South of Brazil, using a multiplex PCR. One hundred and eighteen R. equi isolates were identified by biochemical tests and submitted to a specie-specific and vapA multiplex PCR. These isolates were obtained from: three farms where the R. equi infection has been noticed, two farms where the disease has been not reported and one farm where the disease is...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Rhodococcus equi; Multiplex PCR; Bronchopneumonia; Epidemiology.
Ano: 2008 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1517-83822008000100036
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Molecular detection of adulteration in commercial buffalo meat products by multiplex PCR assay Ciênc. Tecnol. Aliment.
WANG,Lanping; HANG,Xinru; GENG,Rongqing.
Abstract Authentication of commercial buffalo meat products has become a market concern. This study intended to develop and validate a highly species-specific multiplex PCR assay for authentication of buffalo meat products. Four pairs of species-specific primers were used to target mitochondrial cytochrome oxidase I (COI) gene. The assay generated the expected PCR products of 313, 255, 294 and 177 bp for buffalo meat, cattle meat, pork meat and duck meat, respectively. The multiplex PCR assay was sensitive enough to detect 1 pg pure DNA and 0.1% (w/w) adulterated meat under mixed matrices. Market survey revealed about 35.3% of buffalo meat products are adulterated with cattle meat, pork meat or duck meat in China. The adulteration was found in all food...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Buffalo meat; Commercial fraud; Adulteration; Multiplex PCR; Cytochrome oxidase I gene.
Ano: 2019 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0101-20612019000200344
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Molecular diagnosis of Eimeria species affecting naturally infected Gallus gallus. Repositório Alice
CARVALHO, F. S.; WENCESLAU, A. A.; TEIXEIRA, M.; ALBUQUERQUE, G. R..
Abstract - We used PCR to test various protocols and define a technique for DNA extraction directly from chicken-shed stool samples for the identification of Eimeria species that parasitize birds. It was possible to extract and amplify DNA of seven Eimeria species from field stool samples, using both protocols tested; extractions made with phenol/chloroform protocols gave the best results. The primers were specific and sensitive, allowing amplification of samples containing as few as 20 oocysts, both in individual and in a multiplex PCR. Individualized PCR with the phenol/chloroform DNA extraction protocol detected a larger number of Eimeria species. Molecular diagnosis was found to be practical and precise, and can be used for monitoring and...
Tipo: Artigo em periódico indexado (ALICE) Palavras-chave: PCR; Reação em cadeia; Frango; Eimeria; Prevalência; Polimerase; Eimeriosis; Multiplex PCR; Broilers.
Ano: 2011 URL: http://www.alice.cnptia.embrapa.br/alice/handle/doc/908376
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Molecular screening of bovine raw milk for the presence of Shiga toxin-producing Escherichia coli (STEC) on dairy farms Ciênc. Tecnol. Aliment.
Vendramin,Tatiane; Kich,Débora Mara; Molina,Rachel Dias; Souza,Claucia Fernanda Volken de; Salvatori,Rosangela Uhrig; Pozzobon,Adriane; Bustamante-filho,Ivan Cunha.
Milkborne transmission of Shiga toxin- producing Escherichia coli (STEC) has raised considerable concern due to recent outbreaks worldwide and poses a threat to public health. The aim of this study was to develop a sensitive and specific multiplex PCR assay to detect the presence of STEC in bovine raw milk. To identify E. coli (ATCC 25922) contamination, the gene uspA was used, and PCR sensitivity and specificity were accessed by testing diluted samples ranging from 2 to 2.0 × 10(6) CFU/mL. To detect STEC, the stx1 and stx2 genes were selected as targets. After reaction standardization, the multiplex assay was tested in raw milk collected from 101 cows on dairy farms. PCR assay for E. coli detection had a specificity of 100% and sensitivity of 79%...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Shiga-toxin; Cattle; Multiplex PCR; Milk.
Ano: 2014 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0101-20612014000300025
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Occurrence and characteristics of virulence genes of Escherichia coli strains isolated from healthy dairy cows in Inner Mongolia, China BJM
Huasai,Simujide; Chen,Aorigele; Wang,Chun-jie; Li,Yu; Tongrige,Bai.
Virulence genes of Escherichia coli (E. coli) isolates from healthy dairy cows were identified and characterized by a multiplex PCR assay and serogrouping test. The results showed that among the target genes, eaeA was most frequently detected, accounting for 22.11% (67/303) in all strains from 101 cows. For categorization of E. coli, aEPEC was the category with widest distribution detected in 55 (18.15%) strains from 22 cattle. All of 84 PCR-positive strains belonged to 14 O serogroups, and O149 (25.00%) was most common identified, followed by O2 (17.86%), O8 (11.90%) and O103 (9.52%) with relatively high prevalence.
Tipo: Info:eu-repo/semantics/article Palavras-chave: E. coli; Virulence gene; Multiplex PCR; Serogroup; Healthy dairy cow.
Ano: 2012 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1517-83822012000200013
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Ocurrence of Staphylococcus aureus and multiplex pcr detection of classic enterotoxin genes in cheese and meat products BJM
Pelisser,Marcia Regina; Klein,Cátia Silene; Ascoli,Kelen Regina; Zotti,Thaís Regina; Arisi,Ana Carolina Maisonnave.
Multiplex PCR was used to investigate the presence of enterotoxins genes (sea, seb, sec, sed and see) and femA gene (specific for Staphylococcus aureus) in coagulase-positive staphylococci (CPS) isolated from cheese and meat products. From 102 CPS isolates, 91 were positive for femA, 10 for sea, 12 for sed and four for see.
Tipo: Info:eu-repo/semantics/article Palavras-chave: Staphylococcus aureus; Enterotoxins; Multiplex PCR.
Ano: 2009 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1517-83822009000100025
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Preimplantation genetic diagnosis of Von Hippel-Lindau disease cancer syndrome by combined mutation and segregation analysis Genet. Mol. Biol.
Sumita,Denilce R.; Rocha,José Cláudio C.; Iaconelli Jr.,Assumpto; Borges Jr.,Edson; Pereira,Lygia V..
Von Hippel-Lindau (VHL) disease is an autosomal dominant cancer syndrome, associated with the development of tumors and cysts in multiple organ systems, whose expression and age of onset are highly variable. The VHL disease tumor suppressor gene (VHL) maps to 3p25-p26 and mutations ranging from a single base change to large deletions have been detected in patients with VHL disease. We developed a single cell PCR protocol for preimplantation genetic diagnosis (PGD) of VHL disease to select unaffected embryos on the basis of the detection of the specific mutation and segregation analysis of polymorphic linked markers. Multiplex-nested PCR using single buccal cells of an affected individual were performed in order to test the accuracy and reliability of this...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Multiplex PCR; Preimplantation genetic diagnosis; Von Hippel-Lindau disease; VHL gene.
Ano: 2007 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1415-47572007000300005
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Presence of enterotoxigenic Staphylococcus aureus in artisan fruit salads in the city of San Luis, Argentina BJM
Lucero Estrada,Cecilia S.M.; Alcaráz,Lucia E.; Satorres,Sara E.; Manfredi,Eduardo; Velázquez,Lidia del C..
An increase in the consumption of fruit juices and minimally processed fruits salads has been observed in recent years all over the world. In this work, the microbiological quality of artisan fruit salads was analysed. Faecal coliforms, Salmonella spp, Shigella spp, Yersinia enterocolitica and Escherichia coli O157:H7 were not detected; nevertheless, eleven strains of Staphylococcus aureus were isolated. By multiplex PCR, all isolates showed positive results for S. aureus 16S rRNA gene and 63.6% of them were positive for sea gene. Furthermore, PCR sea positive strains were able to produce the corresponding enterotoxin. Finally, the inactivation of these strains in fruit salads by nisin, lysozyme and EDTA, was studied. EDTA produced a total S. aureus growth...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Staphylococcus aureus; Enterotoxigenic strains; Fruit salads; Antimicrobial compounds; Multiplex PCR.
Ano: 2013 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1517-83822013000400018
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