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J Clin Microbiol. 2018 Oct 25;56(11). pii: e00940-18. doi: 10.1128/JCM.00940-18. Print 2018 Nov.

Genotyping and Quantifying Lyme Pathogen Strains by Deep Sequencing of the Outer Surface Protein C (ospC) Locus.

Author information

1
Department of Biological Sciences, Hunter College of the City University of New York, New York, New York, USA.
2
Graduate Center of the City University of New York, New York, New York, USA.
3
Division of Vector Control, Suffolk County Department of Public Works, Yaphank, New York, USA.
4
Department of Physiology and Biophysics, Institute for Computational Biomedicine, Weil Cornell Medical College, New York, New York, USA.
5
Department of Biological Sciences, Hunter College of the City University of New York, New York, New York, USA weigang@genectr.hunter.cuny.edu.

Abstract

A mixed infection of a single tick or host by Lyme disease spirochetes is common and a unique challenge for the diagnosis, treatment, and surveillance of Lyme disease. Here, we describe a novel protocol for differentiating Lyme strains on the basis of deep sequencing of the hypervariable outer surface protein C locus (ospC). Improving upon the traditional DNA-DNA hybridization method, the next-generation sequencing-based protocol is high throughput, quantitative, and able to detect new pathogen strains. We applied the method to more than one hundred infected Ixodes scapularis ticks collected from New York State, USA, in 2015 and 2016. An analysis of strain distributions within individual ticks suggests an overabundance of multiple infections by five or more strains, inhibitory interactions among coinfecting strains, and the presence of a new strain closely related to Borreliella bissettiae A supporting bioinformatics pipeline has been developed. The newly designed pair of universal ospC primers target intergenic sequences conserved among all known Lyme pathogens. The protocol could be used for culture-free identification and quantification of Lyme pathogens in wildlife and potentially in clinical specimens.

KEYWORDS:

Borrelia burgdorferi; Borreliella; Ixodes scapularis; Lyme disease; frequency-dependent selection; next-generation sequencing; outer surface protein C

PMID:
30158192
PMCID:
PMC6204676
[Available on 2019-04-25]
DOI:
10.1128/JCM.00940-18

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