Detection of Borrelia burgdorferi, Anaplasma phagocytophilum and Babesia microti, with two different multiplex PCR assays

Ticks Tick Borne Dis. 2014 Apr;5(3):349-51. doi: 10.1016/j.ttbdis.2013.12.001. Epub 2014 Jan 18.

Abstract

We have developed 2 real-time multiplex PCR assays for detection of Borrelia burgdorferi, Anaplasma phagocytophilum, and Babesia microti. The efficiency and sensitivity of each multiplex PCR assay was evaluated using field-collected Ixodes scapularis ticks that were positive for each of the pathogens, cloned plasmids harboring each of the PCR targets, and laboratory I. scapularis infected with B. burgdorferi B31. There was no difference in efficiency or sensitivity when comparing the multiplex PCR with the individual PCR reactions. If the 2 multiplex PCR assays are used in the same analysis, field-collected ticks that only harbor B. miyamotoi can also be identified. The multiplex assays are fast and cost-effective methods for screening and detecting pathogens in ticks, when compared to single-target PCR.

Keywords: Multiplex PCR; Pathogens; Ticks.

Publication types

  • Comparative Study
  • Evaluation Study

MeSH terms

  • Anaplasma phagocytophilum / genetics
  • Anaplasma phagocytophilum / isolation & purification*
  • Animals
  • Babesia microti / genetics
  • Babesia microti / isolation & purification*
  • Bacterial Proteins / genetics
  • Borrelia burgdorferi / genetics
  • Borrelia burgdorferi / isolation & purification*
  • DNA Primers / genetics
  • DNA, Bacterial / genetics
  • DNA, Protozoan / genetics
  • Ixodes / parasitology*
  • Multiplex Polymerase Chain Reaction / economics
  • Multiplex Polymerase Chain Reaction / methods*
  • Protozoan Proteins / genetics
  • Real-Time Polymerase Chain Reaction / economics
  • Real-Time Polymerase Chain Reaction / methods
  • Sensitivity and Specificity
  • Time Factors

Substances

  • Bacterial Proteins
  • DNA Primers
  • DNA, Bacterial
  • DNA, Protozoan
  • Protozoan Proteins