Next generation multilocus sequence typing (NGMLST) and the analytical software program MLSTEZ enable efficient, cost-effective, high-throughput, multilocus sequencing typing.
Fungal Genet Biol
; 75: 64-71, 2015 Feb.
Article
em En
| MEDLINE
| ID: mdl-25624069
ABSTRACT
Multilocus sequence typing (MLST) has become the preferred method for genotyping many biological species, and it is especially useful for analyzing haploid eukaryotes. MLST is rigorous, reproducible, and informative, and MLST genotyping has been shown to identify major phylogenetic clades, molecular groups, or subpopulations of a species, as well as individual strains or clones. MLST molecular types often correlate with important phenotypes. Conventional MLST involves the extraction of genomic DNA and the amplification by PCR of several conserved, unlinked gene sequences from a sample of isolates of the taxon under investigation. In some cases, as few as three loci are sufficient to yield definitive results. The amplicons are sequenced, aligned, and compared by phylogenetic methods to distinguish statistically significant differences among individuals and clades. Although MLST is simpler, faster, and less expensive than whole genome sequencing, it is more costly and time-consuming than less reliable genotyping methods (e.g. amplified fragment length polymorphisms). Here, we describe a new MLST method that uses next-generation sequencing, a multiplexing protocol, and appropriate analytical software to provide accurate, rapid, and economical MLST genotyping of 96 or more isolates in single assay. We demonstrate this methodology by genotyping isolates of the well-characterized, human pathogenic yeast Cryptococcus neoformans.
Palavras-chave
Texto completo:
1
Base de dados:
MEDLINE
Assunto principal:
Software
/
Cryptococcus neoformans
/
Tipagem de Sequências Multilocus
/
Sequenciamento de Nucleotídeos em Larga Escala
Tipo de estudo:
Health_economic_evaluation
Limite:
Humans
Idioma:
En
Revista:
Fungal Genet Biol
Assunto da revista:
GENETICA
/
MICROBIOLOGIA
Ano de publicação:
2015
Tipo de documento:
Article