Rapid detection and differentiation of Bartonella spp. by a single-run real-time PCR.
A Real-time PCR for the identification and differentiation of Bartonella spp. based on the detection of mutations in an internal region of the gltA gene by thermal analysis was developed. The assay included a simultaneous detection of the amplicons by direct hybridization of LCRed and fluorescein-labelled probes coupled with melting curve analysis by the use of fluorescence resonance energy transfer technology. The protocol allowed establishing genospecie identity in less than 1 h without a need for restriction enzyme digestion or gel electrophoresis. The method is simple, reproducible, rapid, specific and potentially transferable to clinical samples.
Department of Infectious, Parasitic and Immune-mediated Diseases, Istituto Superiore di Sanità, viale Regina Elena 299, 00161 Rome, Italy. ciervo@iss.it
Alessandra
A
Ciervo
Lorenzo
L
Ciceroni
D001473:Bartonella / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification*; D005798:Genes, Bacterial / Q000235:genetics; D008969:Molecular Sequence Data; D009691:Nucleic Acid Denaturation; D016133:Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D017422:Sequence Analysis, DNA; D013045:Species Specificity; D013997:Time Factors