A LightCycler TaqMan assay for detection of Borrelia burgdorferi sensu lato in clinical samples.
Lyme disease (LD) is an infection caused by an ixodid tick-borne spirochete, Borrelia burgdorferi sensu lato. LD manifests itself as a multisystem inflammatory disease that affects the skin in its early localized stage and spreads to the joints, nervous system, heart, and, to a lesser extent, other organ systems in its later disseminated stages. If diagnosed and treated early with appropriate antibiotics, LD is almost always readily cured. Developing a highly sensitive and specific real-time polymerase chain reaction assay could be very useful in improving the diagnostic accuracy and decreasing turnaround time for results. We report the development of a LightCycler TaqMan assay targeting the OspA gene for clinical detection of B. burgdorferi sensu lato in various types of biologic samples. This assay was validated by testing a variety of clinical samples including cerebrospinal fluid, synovial fluid, skin biopsies, and blood and culture isolates from skin biopsies. The TaqMan testing results were 100% concordant with previously reported results. Reference strains representing isolates from other geographic regions were also successfully amplified. The developed assay is robust, is highly sensitive and specific for B. burgdorferi sensu lato, and is suitable for clinical detection of the bacterium in biologic samples.
Ivacic
Lynn
L
Molecular Diagnostics Genotyping Laboratory, Marshfield Clinic, Marshfield, Wisconsin, WI 54449, USA.
Reed
Kurt D
KD
Mitchell
Paul D
PD
Ghebranious
Nader
N
D000818:Animals; D000954:Antigens, Surface / Q000235:genetics*; D001425:Bacterial Outer Membrane Proteins / Q000235:genetics*; D001428:Bacterial Vaccines / Q000235:genetics*; D015748:Borrelia burgdorferi Group / Q000235:genetics / Q000302:isolation & purification*; D004269:DNA, Bacterial / Q000032:analysis / Q000302:isolation & purification; D006801:Humans; D008074:Lipoproteins / Q000235:genetics*; D008193:Lyme Disease / Q000175:diagnosis / Q000382:microbiology*; D051379:Mice; D016133:Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D019914:Taq Polymerase
D000954:Antigens, Surface; D001425:Bacterial Outer Membrane Proteins; D001428:Bacterial Vaccines; D004269:DNA, Bacterial; D008074:Lipoproteins; C072045:OspA protein; D019914:Taq Polymerase