Lyme Borreliosis--the Utility of Improved Real-Time PCR Assay in the Detection of Borrelia burgdorferi Infections

Adv Clin Exp Med. 2015 Jul-Aug;24(4):663-70. doi: 10.17219/acem/28625.

Abstract

Background: Infections of Borrelia burgdorferi sensu lato reveal clinical manifestations affecting numerous organs and tissues. The standard diagnostic procedure of these infections is quite simple if a positive history of tick exposure or typical erythema migrans appears. Lack of unequivocal clinical symptoms creates the necessity for further evaluation with laboratory tests.

Objectives: This study discusses the utility of a novel, improved, well-optimized, sensitive and highly specific quantitative real-time PCR assay for the diagnostics of infections caused by Borrelia burgdorferi sensu lato.

Material and methods: We designed an improved, specific, highly sensitive real-time quantitative polymerase chain reaction (RQ-PCR) assay for the detection and quantification of all Borrelia burgdorferi genotypes. A wide validation effort was undertaken to ensure confidence in the highly sensitive and specific detection of B. burgdorferi.

Results: Due to high sensitivity and great specificity, as low as 1.6×10² copies of Borrelia per mL of whole blood could be detected. As much as 12 (3%) negative ELISA IgM results, 14 (2.8%) negative results of Line blot IgM, 11 (3.1%) and 7 (2.7%) of negative ELISA IgG and Line blot IgG results, respectively, were positive in real-time PCR.

Conclusions: The data in this study confirms the high positive predictive value of real-time PCR test in the detection of Borrelia infections.

Publication types

  • Comparative Study
  • Validation Study

MeSH terms

  • Antibodies, Bacterial / blood
  • Borrelia burgdorferi / classification
  • Borrelia burgdorferi / genetics*
  • Borrelia burgdorferi / immunology
  • Calibration
  • DNA, Bacterial / genetics*
  • Enzyme-Linked Immunosorbent Assay
  • Genetic Markers
  • Humans
  • Incidence
  • Lyme Disease / blood
  • Lyme Disease / diagnosis*
  • Lyme Disease / epidemiology
  • Lyme Disease / microbiology
  • Poland / epidemiology
  • Predictive Value of Tests
  • RNA, Ribosomal, 16S / genetics*
  • Real-Time Polymerase Chain Reaction* / standards
  • Reference Standards
  • Reproducibility of Results
  • Ribotyping / methods*
  • Ribotyping / standards
  • Serologic Tests

Substances

  • Antibodies, Bacterial
  • DNA, Bacterial
  • Genetic Markers
  • RNA, Ribosomal, 16S