Use of sloppy molecular beacon probes for identification of mycobacterial species.
J Clin Microbiol
; 47(4): 1190-8, 2009 Apr.
Article
en En
| MEDLINE
| ID: mdl-19171684
ABSTRACT
We report here the use of novel "sloppy" molecular beacon probes in homogeneous PCR screening assays in which thermal denaturation of the resulting probe-amplicon hybrids provides a characteristic set of amplicon melting temperature (T(m)) values that identify which species is present in a sample. Sloppy molecular beacons possess relatively long probe sequences, enabling them to form hybrids with amplicons from many different species despite the presence of mismatched base pairs. By using four sloppy molecular beacons, each possessing a different probe sequence and each labeled with a differently colored fluorophore, four different T(m) values can be determined simultaneously. We tested this technique with 27 different species of mycobacteria and found that each species generates a unique, highly reproducible signature that is unaffected by the initial bacterial DNA concentration. Utilizing this general paradigm, screening assays can be designed for the identification of a wide range of species.
Texto completo:
1
Colección:
01-internacional
Banco de datos:
MEDLINE
Asunto principal:
Técnicas Bacteriológicas
/
Técnicas de Sonda Molecular
/
Mycobacterium
Tipo de estudio:
Diagnostic_studies
/
Evaluation_studies
/
Prognostic_studies
/
Screening_studies
Límite:
Humans
Idioma:
En
Revista:
J Clin Microbiol
Año:
2009
Tipo del documento:
Article
País de afiliación:
Estados Unidos