Development of a multiplex real-time polymerase chain reaction for simultaneous detection of enterohemorrhagic Escherichia coli and enteropathogenic Escherichia coli strains. 2010

Melanie Pavlovic, and Ingrid Huber, and Henrike Skala, and Regina Konrad, and Herbert Schmidt, and Andreas Sing, and Ulrich Busch
Bavarian Health and Food Safety Authority, Oberschleissheim, Germany.

A multiplex real-time polymerase chain reaction (PCR) was developed for the simultaneous detection of genes encoding intimin (eae) and all variants of Shiga toxins 1 and 2 (stx1 and stx2) in diagnostic samples. The uidA gene encoding a beta-glucuronidase specific for Escherichia coli and Shigella spp. was included in the multiplex PCR assay as an internal amplification control. The multiplex PCR was tested on 30 E. coli reference strains and 174 diagnostic samples already characterized as harboring stx1, stx2, and eae genes. The multiplex PCR correctly detected the genes in all strains examined. No cross reaction was observed with 68 strains representing other gastrointestinal pathogens, normal gastrointestinal flora, or closely related bacteria, reflecting 100% specificity of the assay. The detection limits of the multiplex PCR were 5 genome equivalents for stx2 and 50 genome equivalents for eae and stx1.

UI MeSH Term Description Entries
D004269 DNA, Bacterial Deoxyribonucleic acid that makes up the genetic material of bacteria. Bacterial DNA
D004926 Escherichia coli A species of gram-negative, facultatively anaerobic, rod-shaped bacteria (GRAM-NEGATIVE FACULTATIVELY ANAEROBIC RODS) commonly found in the lower part of the intestine of warm-blooded animals. It is usually nonpathogenic, but some strains are known to produce DIARRHEA and pyogenic infections. Pathogenic strains (virotypes) are classified by their specific pathogenic mechanisms such as toxins (ENTEROTOXIGENIC ESCHERICHIA COLI), etc. Alkalescens-Dispar Group,Bacillus coli,Bacterium coli,Bacterium coli commune,Diffusely Adherent Escherichia coli,E coli,EAggEC,Enteroaggregative Escherichia coli,Enterococcus coli,Diffusely Adherent E. coli,Enteroaggregative E. coli,Enteroinvasive E. coli,Enteroinvasive Escherichia coli
D004927 Escherichia coli Infections Infections with bacteria of the species ESCHERICHIA COLI. E coli Infections,E. coli Infection,Infections, E coli,Infections, Escherichia coli,E coli Infection,E. coli Infections,Escherichia coli Infection,Infection, E coli,Infection, E. coli,Infection, Escherichia coli
D005243 Feces Excrement from the INTESTINES, containing unabsorbed solids, waste products, secretions, and BACTERIA of the DIGESTIVE SYSTEM.
D005798 Genes, Bacterial The functional hereditary units of BACTERIA. Bacterial Gene,Bacterial Genes,Gene, Bacterial
D006801 Humans Members of the species Homo sapiens. Homo sapiens,Man (Taxonomy),Human,Man, Modern,Modern Man
D012680 Sensitivity and Specificity Binary classification measures to assess test results. Sensitivity or recall rate is the proportion of true positives. Specificity is the probability of correctly determining the absence of a condition. (From Last, Dictionary of Epidemiology, 2d ed) Specificity,Sensitivity,Specificity and Sensitivity
D014644 Genetic Variation Genotypic differences observed among individuals in a population. Genetic Diversity,Variation, Genetic,Diversity, Genetic,Diversities, Genetic,Genetic Diversities,Genetic Variations,Variations, Genetic
D014774 Virulence The degree of pathogenicity within a group or species of microorganisms or viruses as indicated by case fatality rates and/or the ability of the organism to invade the tissues of the host. The pathogenic capacity of an organism is determined by its VIRULENCE FACTORS. Pathogenicity
D016133 Polymerase Chain Reaction In vitro method for producing large amounts of specific DNA or RNA fragments of defined length and sequence from small amounts of short oligonucleotide flanking sequences (primers). The essential steps include thermal denaturation of the double-stranded target molecules, annealing of the primers to their complementary sequences, and extension of the annealed primers by enzymatic synthesis with DNA polymerase. The reaction is efficient, specific, and extremely sensitive. Uses for the reaction include disease diagnosis, detection of difficult-to-isolate pathogens, mutation analysis, genetic testing, DNA sequencing, and analyzing evolutionary relationships. Anchored PCR,Inverse PCR,Nested PCR,PCR,Anchored Polymerase Chain Reaction,Inverse Polymerase Chain Reaction,Nested Polymerase Chain Reaction,PCR, Anchored,PCR, Inverse,PCR, Nested,Polymerase Chain Reactions,Reaction, Polymerase Chain,Reactions, Polymerase Chain

Related Publications

Melanie Pavlovic, and Ingrid Huber, and Henrike Skala, and Regina Konrad, and Herbert Schmidt, and Andreas Sing, and Ulrich Busch
June 2010, Foodborne pathogens and disease,
Melanie Pavlovic, and Ingrid Huber, and Henrike Skala, and Regina Konrad, and Herbert Schmidt, and Andreas Sing, and Ulrich Busch
January 2012, Frontiers in cellular and infection microbiology,
Melanie Pavlovic, and Ingrid Huber, and Henrike Skala, and Regina Konrad, and Herbert Schmidt, and Andreas Sing, and Ulrich Busch
January 2009, Methods in molecular biology (Clifton, N.J.),
Melanie Pavlovic, and Ingrid Huber, and Henrike Skala, and Regina Konrad, and Herbert Schmidt, and Andreas Sing, and Ulrich Busch
July 2018, Journal of food science and technology,
Melanie Pavlovic, and Ingrid Huber, and Henrike Skala, and Regina Konrad, and Herbert Schmidt, and Andreas Sing, and Ulrich Busch
July 2001, Journal of veterinary diagnostic investigation : official publication of the American Association of Veterinary Laboratory Diagnosticians, Inc,
Melanie Pavlovic, and Ingrid Huber, and Henrike Skala, and Regina Konrad, and Herbert Schmidt, and Andreas Sing, and Ulrich Busch
January 2005, Journal of applied microbiology,
Melanie Pavlovic, and Ingrid Huber, and Henrike Skala, and Regina Konrad, and Herbert Schmidt, and Andreas Sing, and Ulrich Busch
May 2009, Diagnostic microbiology and infectious disease,
Melanie Pavlovic, and Ingrid Huber, and Henrike Skala, and Regina Konrad, and Herbert Schmidt, and Andreas Sing, and Ulrich Busch
December 2011, Clinical infectious diseases : an official publication of the Infectious Diseases Society of America,
Melanie Pavlovic, and Ingrid Huber, and Henrike Skala, and Regina Konrad, and Herbert Schmidt, and Andreas Sing, and Ulrich Busch
June 2010, Foodborne pathogens and disease,
Melanie Pavlovic, and Ingrid Huber, and Henrike Skala, and Regina Konrad, and Herbert Schmidt, and Andreas Sing, and Ulrich Busch
February 1997, Research in microbiology,
Copied contents to your clipboard!