Next Article in Journal
Mechanism and Kinetic Analysis of Degradation of Atrazine by US/PMS
Previous Article in Journal
Dietary and Activity Factors Influence Poor Sleep and the Sleep-Obesity Nexus among Children
Previous Article in Special Issue
A Review on Equine Piroplasmosis: Epidemiology, Vector Ecology, Risk Factors, Host Immunity, Diagnosis and Control
Article Menu
Issue 10 (May-2) cover image

Export Article

Open AccessArticle

Single Core Genome Sequencing for Detection of both Borrelia burgdorferi Sensu Lato and Relapsing Fever Borrelia Species

1
Milford Molecular Diagnostics, Milford, CT 06460, USA
2
School of Biological, Earth and Environmental Sciences, University College Cork, T23 N73K Cork, Ireland
3
Department of medicine, University College Dublin, D04 V1W8 Dublin, Ireland
4
Mater Misericordiae University Hospital, D07 R2WY Dublin, Ireland
*
Author to whom correspondence should be addressed.
Int. J. Environ. Res. Public Health 2019, 16(10), 1779; https://doi.org/10.3390/ijerph16101779
Received: 30 January 2019 / Revised: 20 February 2019 / Accepted: 16 May 2019 / Published: 20 May 2019
(This article belongs to the Special Issue Ticks and Tick Vectored Diseases—Biology to Society)
  |  
PDF [3122 KB, uploaded 20 May 2019]
  |     |  

Abstract

Lyme disease, initially described as Lyme arthritis, was reported before nucleic-acid based detection technologies were available. The most widely used diagnostic tests for Lyme disease are based on the serologic detection of antibodies produced against antigens derived from a single strain of Borrelia burgdorferi. The poor diagnostic accuracy of serological tests early in the infection process has been noted most recently in the 2018 Report to Congress issued by the U.S. Department of Health and Human Services Tick-Borne Disease Working Group. Clinical Lyme disease may be caused by a diversity of borreliae, including those classified as relapsing fever species, in the United States and in Europe. It is widely accepted that antibiotic treatment of Lyme disease is most successful during this critical early stage of infection. While genomic sequencing is recognized as an irrefutable direct detection method for laboratory diagnosis of Lyme borreliosis, development of a molecular diagnostic tool for all clinical forms of borreliosis is challenging because a “core genome” shared by all pathogenic borreliae has not yet been identified. After a diligent search of the GenBank database, we identified two highly conserved segments of DNA sequence among the borrelial 16S rRNA genes. We further developed a pair of Borrelia genus-specific PCR primers for amplification of a segment of borrelial 16S rRNA gene as a “core genome” to be used as the template for routine Sanger sequencing-based metagenomic direct detection test. This study presented examples of base-calling DNA sequencing electropherograms routinely generated in a clinical diagnostic laboratory on DNA extracts of human blood specimens and ticks collected from human skin bites and from the environment. Since some of the tick samples tested were collected in Ireland, borrelial species or strains not known to exist in the United States were also detected by analysis of this 16S rRNA “core genome”. We recommend that hospital laboratories located in Lyme disease endemic areas begin to use a “core genome” sequencing test to routinely diagnose spirochetemia caused by various species of borreliae for timely management of patients at the early stage of infection. View Full-Text
Keywords: Lyme disease; core genome; DNA sequencing; borreliosis; relapsing fever borreliae; metagenomic diagnosis; Borrelia; genus-specific PCR primers; second PCR; direct detection; Sanger sequencing Lyme disease; core genome; DNA sequencing; borreliosis; relapsing fever borreliae; metagenomic diagnosis; Borrelia; genus-specific PCR primers; second PCR; direct detection; Sanger sequencing
Figures

Figure 1

This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited (CC BY 4.0).

Supplementary material

SciFeed

Share & Cite This Article

MDPI and ACS Style

Lee, S.H.; Healy, J.E.; Lambert, J.S. Single Core Genome Sequencing for Detection of both Borrelia burgdorferi Sensu Lato and Relapsing Fever Borrelia Species. Int. J. Environ. Res. Public Health 2019, 16, 1779.

Show more citation formats Show less citations formats

Note that from the first issue of 2016, MDPI journals use article numbers instead of page numbers. See further details here.

Related Articles

Article Metrics

Article Access Statistics

1

Comments

[Return to top]
Int. J. Environ. Res. Public Health EISSN 1660-4601 Published by MDPI AG, Basel, Switzerland RSS E-Mail Table of Contents Alert
Back to Top