FRET Probe System for Multiplex qPCR Detection of Species.

BACKGROUND

Laboratory diagnosis of Lyme borreliosis refers to some methods with known limitations. Molecular diagnostics using specific nucleic acid probes may overcome some of these limitations.

METHODS

We describe the novel reporter fluorescence real-time polymerase chain reaction (PCR) probe system for detection of species. Advantages of the system include having cheap conventional fluorescence dyes, easy primer design, no restrictions for PCR product lengths, robustness, high sequence specificity, applicability for multiplex real-time PCRs, melting curve analysis (single nucleotide polymorphism analysis) over a large temperature range, high sensitivity, and easy adaptation of conventional PCRs.

RESULTS

Using the probe system we were able to detect all nine tested European species belonging to the (sensu lato) complex and differentiated them from relapsing fever species. As few as 10 copies of in one PCR reaction were detectable.

CONCLUSION

We established a novel multiplex probe real-time PCR system, designated , that is simple, robust, and incorporates melting curve analysis for the detection and in the differentiation of European species belonging to the s.l. complex.