Isolation and detection of Borrelia burgdorferi DNA from cerebral spinal fluid, synovial fluid, blood, urine, and ticks using the Roche MagNA Pure system and real-time PCR.
The Roche MagNA Pure automated nucleic acid extraction system was tested for its ability to extract Borrelia burgdorferi DNA from a diverse set of spiked specimen types including blood, cerebral spinal fluid, synovial fluid, urine and ticks. A method comparison between MagNA Pure automated extraction and manual extraction, using either QIAamp columns or phenol/chloroform extraction, showed equivalent detection sensitivities for all methodologies with all specimen types (except for urine, in which case QIAamp extraction was twofold less sensitive). Eighty positive clinical specimens (as determined by an independent testing method), including 76 synovial fluid, and 4 cerebral spinal fluid specimens, were found to be positive by the MagNA Pure/real-time PCR method of extraction and detection. This data shows that the MagNA Pure system can be used to extract B. burgdorferi DNA from clinical specimens, and when combined with real-time PCR, the result is an extremely sensitive assay with limited hands on time and rapid turn around times.
Exner
Maurice M
MM
Quest Diagnostics' Nichols Institute, San Juan, Capistrano, CA, USA. exnerm@questdiagnostics.com
Lewinski
Michael A
MA
D000818:Animals; D001431:Bacteriological Techniques; D001483:Base Sequence; D025065:Borrelia burgdorferi / Q000302:isolation & purification*; D004269:DNA, Bacterial / Q000302:isolation & purification*; D005260:Female; D006801:Humans; D008193:Lyme Disease / Q000097:blood / Q000134:cerebrospinal fluid / Q000652:urine; D008297:Male; D008969:Molecular Sequence Data; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D012494:Sampling Studies; D012680:Sensitivity and Specificity; D013582:Synovial Fluid / Q000382:microbiology*; D013987:Ticks / Q000382:microbiology*
D004269:DNA, Bacterial