Optimization of polymerase chain reaction for the detection of Borrelia burgdorferi in biologic specimens.
This study describes the use of a newly constructed set of primers that amplifies an 85-base pair (bp) segment of Borrelia burgdorferi chromosomal DNA. This 85-bp product is not produced when other Borrelia species, Leptospira, or other bacteria are subjected to polymerase chain reaction (PCR). We also describe a rapid method of optimizing the amplification of B. burgdorferi DNA from canine ethylenediaminetetraacetic acid-treated blood and urine samples that circumvents some of the problems encountered due to low number of spirochetes in clinical specimens and that removes inhibiting substances, which improves the PCR diagnosis of canine Lyme borreliosis.
Kaufman
A C
AC
Department of Small Animal Medicine, College of Veterinary Medicine, University of Georgia, Athens 30602.
Greene
C E
CE
McGraw
R A
RA
D000818:Animals; D001483:Base Sequence; D015748:Borrelia burgdorferi Group / Q000235:genetics / Q000302:isolation & purification*; D002876:Chromosomes, Bacterial; D017931:DNA Primers; D004269:DNA, Bacterial / Q000032:analysis; D004283:Dog Diseases; D004285:Dogs; D007921:Leptospira interrogans / Q000235:genetics / Q000302:isolation & purification; D008193:Lyme Disease / Q000175:diagnosis / Q000662:veterinary*; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods / Q000662:veterinary*; D012680:Sensitivity and Specificity
D017931:DNA Primers; D004269:DNA, Bacterial