Detection and differentiation of lyme spirochetes and other tick-borne pathogens from blood using real-time PCR with molecular beacons

Samantha Schlachter, Kamfai Chan, Salvatore A.E. Marras, Nikhat Parveen

Research output: Chapter in Book/Report/Conference proceedingChapter

6 Scopus citations

Abstract

Real-time PCR assays have recently been implemented in diagnostics for many bacterial pathogens, allowing rapid and accurate detection, which ultimately results in improved clinical intervention. Here, we describe a sensitive method of detection for three common tick-borne pathogens Borrelia burgdorferi, Anaplasma phagocytophilum, and Babesia microti since coinfections with these pathogens have started occurring with increasing frequency over the last several years in both North America and Europe. A shared geographic region, the same tick vectors, and similar transmission cycle all favor simultaneous transmission of these three tick-borne pathogens. Furthermore, early symptoms of the diseases are often similar and somewhat nonspecific leading to poor clinical identification. The multiplex real-time PCR assay we describe here utilizes gene-specific primers, molecular beacon probes tagged with different fluorophores, and optimized PCR conditions to detect even small amounts of specific pathogen DNA without interference. Application of this detection method will offer better diagnostics for acute and persistent infection compared to the two-tier serological tests that are currently approved in North America and Europe, which do not necessarily detect active infection.

Original languageEnglish (US)
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages155-170
Number of pages16
DOIs
StatePublished - 2017

Publication series

NameMethods in Molecular Biology
Volume1616

All Science Journal Classification (ASJC) codes

  • Genetics
  • Molecular Biology

Keywords

  • Anaplasma phagocytophilium
  • Anaplasmosis
  • Babesia microti
  • Babesiosis
  • Borrelia burgdorferi
  • Lyme disease
  • Molecular beacons
  • Mutliplex assay
  • Real-time PCR
  • Tick-borne pathogens

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