A Collagenase-Targeted Multiplex PCR Assay for Identification of Vibrio alginolyticus, Vibrio cholerae, and Vibrio parahaemolyticus

2005 ◽  
Vol 68 (1) ◽  
pp. 150-153 ◽  
Author(s):  
ANGELA DI PINTO ◽  
GIUSEPPINA CICCARESE ◽  
GIUSEPPINA TANTILLO ◽  
DOMENICO CATALANO ◽  
VITO TONY FORTE
Keyword(s):  
Multiplex Pcr ◽  
Vibrio Cholerae ◽  
Vibrio Parahaemolyticus ◽  
Food Samples ◽  
Vibrio Alginolyticus ◽  
Pcr Assay ◽  
Molecular Approaches ◽  
Multiplex Pcr Assay ◽  
Primer Sets ◽  
Species Specific

A multiplex PCR assay using three collagenase-targeted primer pairs for the species-specific detection of Vibrio alginolyticus, Vibrio cholerae, and Vibrio parahaemolyticus was developed. The results highlight the species specificity of the three primer sets designed. Because of the increasing importance of Vibrio spp. in human foodborne diseases, molecular approaches for routine microbial screening and monitoring of clinical, environmental, and food samples also have become more important. The results of this study indicate that the gene coding for collagenase should be used as an alternative molecular target to discriminate among the three Vibrio species.

Multiplex PCR assay for identification of three major pathogenic Vibrio spp., Vibrio cholerae, Vibrio parahaemolyticus, and Vibrio vulnificus

2011 ◽  
Vol 25 (4) ◽  
pp. 174-176 ◽  
Author(s):  
Hidemasa Izumiya ◽  
Kazutoshi Matsumoto ◽  
Shunsuke Yahiro ◽  
Jiyoung Lee ◽  
Masatomo Morita ◽  
...  
Keyword(s):  
Multiplex Pcr ◽  
Vibrio Cholerae ◽  
Vibrio Parahaemolyticus ◽  
Vibrio Vulnificus ◽  
Pcr Assay ◽  
Multiplex Pcr Assay ◽  
Pathogenic Vibrio ◽  
Vibrio Spp

Multiplex PCR Detection of Enterotoxin Genes in Aeromonas spp. from Suspect Food Samples in Northern Taiwan

2008 ◽  
Vol 71 (10) ◽  
pp. 2094-2099 ◽  
Author(s):  
YU-CHANG CHANG ◽  
JAN-YI WANG ◽  
AMMAIYAPPAN SELVAM ◽  
SHU-CHEN KAO ◽  
SHANG-SHYNG YANG ◽  
...  
Keyword(s):  
Multiplex Pcr ◽  
Diarrheal Disease ◽  
Simultaneous Detection ◽  
Food Poisoning ◽  
Food Samples ◽  
Pcr Assay ◽  
Multiplex Pcr Assay ◽  
Enterotoxin Genes ◽  
Causative Agents ◽  
Northern Taiwan

Aeromonads possess an array of virulence factors and are causative agents of a number of human infections. Among them, genes of one cytotoxic (Act) and two cytotonic (Alt, Ast) enterotoxins are implicated in a human diarrheal disease. A rapid, specific, simultaneous detection of these enterotoxin genes in suspected food poisoning samples is not yet reported. Hence, a multiplex PCR assay was designed to amplify the cytotoxic (act), heat-labile cytotonic (alt), and heat-stable cytotonic (ast) enterotoxin genes of aeromonads. The PCR assay was tested with 133 Aeromonas spp. isolated from suspect food poisoning samples and retail samples of poultry and fish from wet markets in and around Taipei, Northern Taiwan. The Aeromonas spp. isolates were divided into six genotypes based on absence or presence of one or more enterotoxin genes. Of these 133 isolates, Aeromonas caviae (52.5%) and Aeromonas hydrophila (43.4%) were the most frequently isolated species from food poisoning samples and retail samples, respectively. Among the species, A. hydrophila had a significantly higher proportion for harboring three enterotoxin genes than had the others, whereas Aeromonas encheleia, considered a nonpathogen, was found harboring three enterotoxin genes. The multiplex PCR assays are rapid and specific, and provide a useful tool for the detection and genotyping of enterotoxin genes of aeromonads.

scholarly journals Species-Specific Identification of Human Adenoviruses by a Multiplex PCR Assay

2000 ◽  
Vol 38 (11) ◽  
pp. 4114-4120 ◽  
Author(s):  
WanHong Xu ◽  
Mike C. McDonough ◽  
Dean D. Erdman
Keyword(s):  
Multiplex Pcr ◽  
Human Adenovirus ◽  
Cost Effective ◽  
Pcr Assay ◽  
Multiplex Pcr Assay ◽  
Amplicon Size ◽  
Species Specific ◽  
Fiber Gene ◽  
Amplification Reaction

A multiplex PCR assay was developed by using primers to the fiber gene that could differentiate human adenovirus (Ad) species A through F in a single amplification reaction. The assay correctly identified the species of all 49 recognized Ad prototype strains as well as 180 geographically and temporally diverse Ad field isolates. Ad serotype 6 (Ad6) (species C), Ad16 (species B), Ad31 (species A), and Ad40 and Ad41 (species F) could also be distinguished by amplicon size within each respective species. In comparison, a previously described Ad species-specific multiplex PCR assay that used primers to the Ad hexon gene gave equivocal results with several serotypes of species B, whereas our multiplex assay amplified all species B serotypes equally well. Our multiplex PCR assay will permit rapid, accurate, and cost-effective classification of Ad isolates.

scholarly journals Multiplex PCR Assay for Detection and Identification of Clostridium botulinum Types A, B, E, and F in Food and Fecal Material

2001 ◽  
Vol 67 (12) ◽  
pp. 5694-5699 ◽  
Author(s):  
Miia Lindström ◽  
Riikka Keto ◽  
Annukka Markkula ◽  
Mari Nevas ◽  
Sebastian Hielm ◽  
...  
Keyword(s):  
Multiplex Pcr ◽  
Clostridium Botulinum ◽  
Botulinum Neurotoxin ◽  
Bacterial Species ◽  
Type A ◽  
Food Samples ◽  
Type B ◽  
Fecal Material ◽  
Pcr Assay ◽  
Multiplex Pcr Assay

ABSTRACT Botulism is diagnosed by detecting botulinum neurotoxin andClostridium botulinum cells in the patient and in suspected food samples. In this study, a multiplex PCR assay for the detection of Clostridium botulinum types A, B, E, and F in food and fecal material was developed. The method employs four new primer pairs with equal melting temperatures, each being specific to botulinum neurotoxin gene type A, B, E, or F, and enables a simultaneous detection of the four serotypes. A total of 43 C. botulinum strains and 18 strains of other bacterial species were tested. DNA amplification fragments of 782 bp for C. botulinum type A alone, 205 bp for type B alone, 389 bp for type E alone, and 543 bp for type F alone were obtained. Other bacterial species, including C. sporogenes and the nontoxigenic nonproteolytic C. botulinum-like organisms, did not yield a PCR product. Sensitivity of the PCR for types A, E, and F was 102 cells and for type B was 10 cells per reaction mixture. With a two-step enrichment, the detection limit in food and fecal samples varied from 10−2 spore/g for types A, B, and F to 10−1 spore/g of sample material for type E. Of 72 natural food samples investigated, two were shown to contain C. botulinum type A, two contained type B, and one contained type E. The assay is sensitive and specific and provides a marked improvement in the PCR diagnostics of C. botulinum.

scholarly journals cpn60 Gene-Based Multiplex-PCR Assay for Simultaneous Identification of Streptococcal Species

2006 ◽  
Vol 75 (2) ◽  
pp. 235-240 ◽  
Author(s):  
A. Dmitriev ◽  
M. Bhide ◽  
I. Mikula
Keyword(s):  
Multiplex Pcr ◽  
Streptococcus Agalactiae ◽  
Streptococcus Dysgalactiae ◽  
Pcr Assay ◽  
Streptococcus Uberis ◽  
Multiplex Pcr Assay ◽  
Streptococcal Species ◽  
Species Specific ◽  
Simultaneous Identification

The cpn60 genes of Streptococcus agalactiae, Streptococcus dysgalactiae and Streptococcus uberis were sequenced and a certain polymorphism of cpn60 genes was revealed. Presumable species-specific pairs of primers were designed and their specificity was confirmed by PCR. Based on these data, the cpn60 gene-based multiplex-PCR assay was developed. It was found to be effective for simultaneous identification of S. agalactiae, S. dysgalactiae and S. uberis strains.

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