Evaluation of multiplex tandem PCR (MT-PCR) assays for the detection of bacterial resistance genes among Enterobacteriaceae in clinical urines.
J Antimicrob Chemother
; 74(2): 349-356, 2019 02 01.
Article
en En
| MEDLINE
| ID: mdl-30476137
ABSTRACT
Background:
Increasing resistance drives empirical use of less potent and previously reserved antibiotics, including for urinary tract infections (UTIs). Molecular profiling, without culture, might better guide early therapy.Objectives:
To explore the potential of AusDiagnostics multiplex tandem (MT) PCR UTI assays.Methods:
Two MT-PCR assays were developed successively, seeking 8 or 16 resistance genes. Amplification was tracked in real time, with melting temperatures used to confirm product identity. Assays were variously performed on (i) extracted DNA; (ii) cultured bacteria; (iii) urine spiked with reference strains; and (iv) bacteria harvested from clinical urines. Results were compared with those from sequencing, real-time SybrGreen PCR or phenotypic susceptibility.Results:
Performance was similar irrespective of whether DNA, cultures or urines were used, with >90% sensitivity and specificity with respect to common ß-lactamases, dfr genes and aminoglycoside resistance determinants except aadA1/A2/A3, for which carriage correlated poorly with streptomycin resistance. Fluoroquinolone-susceptible and -resistant Escherichia coli (but not other species) were distinguished by the melting temperatures of their gyrA PCR products. The time from urine to results was <3âh.Conclusions:
The MT-PCR assays rapidly identified resistance genes from Gram-negative bacteria in urines as well as from cultivated bacteria. Used directly on urines, this assay has the potential to guide early therapy.
Texto completo:
1
Banco de datos:
MEDLINE
Asunto principal:
Farmacorresistencia Bacteriana Múltiple
/
Enterobacteriaceae
/
Infecciones por Enterobacteriaceae
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Reacción en Cadena de la Polimerasa Multiplex
/
Antibacterianos
Tipo de estudio:
Diagnostic_studies
/
Evaluation_studies
/
Prognostic_studies
Límite:
Humans
Idioma:
En
Revista:
J Antimicrob Chemother
Año:
2019
Tipo del documento:
Article
País de afiliación:
Reino Unido