High-resolution genotyping of Campylobacter species by use of PCR and high-throughput mass spectrometry.
J Clin Microbiol
; 46(4): 1220-5, 2008 Apr.
Article
em En
| MEDLINE
| ID: mdl-18272701
ABSTRACT
In this work we report on a high-throughput mass spectrometry-based technique for the rapid high-resolution identification of Campylobacter jejuni strain types. This method readily distinguishes C. jejuni from C. coli, has a resolving power comparable to that of multilocus sequence typing (MLST), is applicable to mixtures, and is highly automated. The strain typing approach is based on high-performance mass spectrometry, which "weighs" PCR amplicons with enough mass accuracy to unambiguously determine the base composition of each amplicon (i.e., the numbers of A's, G's, C's, and T's). Amplicons are derived from PCR primers which amplify short (<140-bp) regions of the housekeeping genes used by conventional MLST strategies. The results obtained with a challenge panel that comprised 25 strain types of C. jejuni and 25 strain types of C. coli are presented. These samples were parsed and resolved with demonstrated sensitivity down to 10 genomes/PCR from pure isolates.
Texto completo:
1
Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Espectrometria de Massas
/
Campylobacter
/
Reação em Cadeia da Polimerase
Tipo de estudo:
Evaluation_studies
Limite:
Animals
/
Humans
Idioma:
En
Revista:
J Clin Microbiol
Ano de publicação:
2008
Tipo de documento:
Article
País de afiliação:
Estados Unidos