Single-nucleotide polymorphism discrimination using high-resolution melting analysis for the genotyping of Bacillus anthracis. 2015

Sylviane Derzelle
Bacterial Zoonosis Unit, Animal Health Laboratory, University Paris-Est, Anses, 23 Avenue du Général de Gaulle, 94706, Maisons-Alfort Cedex, France, sylviane.derzelle@anses.fr.

High-resolution melting (HRM) is a post-PCR technique that determines with high precision the melt profile of PCR products using a new generation of double-stranded DNA-binding dyes and accurate fluorescence data acquisition over small temperature increments. The method can be used to interrogate small sets of single-nucleotide polymorphisms (SNPs). Here, we describe a simple and cost-effective HRM-based method for the screening of 14 phylogenetically informative SNPs within the genome of Bacillus anthracis that subtype the species into 13 major sublineages or subgroups. Fourteen monoplex and seven duplex SNP-discrimination assays have been designed. We detail the parameters most important for the successful application of HRM for B. anthracis genotyping.

UI MeSH Term Description Entries
D005838 Genotype The genetic constitution of the individual, comprising the ALLELES present at each GENETIC LOCUS. Genogroup,Genogroups,Genotypes
D000818 Animals Unicellular or multicellular, heterotrophic organisms, that have sensation and the power of voluntary movement. Under the older five kingdom paradigm, Animalia was one of the kingdoms. Under the modern three domain model, Animalia represents one of the many groups in the domain EUKARYOTA. Animal,Metazoa,Animalia
D000820 Animal Diseases Diseases that occur in VERTEBRATE animals. Diseases, Animal
D000881 Anthrax An acute infection caused by the spore-forming bacteria BACILLUS ANTHRACIS. It commonly affects hoofed animals such as sheep and goats. Infection in humans often involves the skin (cutaneous anthrax), the lungs (inhalation anthrax), or the gastrointestinal tract. Anthrax is not contagious and can be treated with antibiotics. Bacillus anthracis Infection,Bacillus anthracis Infections
D001408 Bacillus anthracis A species of bacteria that causes ANTHRAX in humans and animals.
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
D044366 Transition Temperature The temperature at which a substance changes from one state or conformation of matter to another. Temperature, Transition,Boiling Point Temperature,Freezing Point Temperature,Melting Point Temperature,Boiling Point Temperatures,Freezing Point Temperatures,Melting Point Temperatures,Temperature, Boiling Point,Temperature, Freezing Point,Temperature, Melting Point,Temperatures, Boiling Point,Temperatures, Freezing Point,Temperatures, Melting Point,Temperatures, Transition,Transition Temperatures
D020641 Polymorphism, Single Nucleotide A single nucleotide variation in a genetic sequence that occurs at appreciable frequency in the population. SNPs,Single Nucleotide Polymorphism,Nucleotide Polymorphism, Single,Nucleotide Polymorphisms, Single,Polymorphisms, Single Nucleotide,Single Nucleotide Polymorphisms

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