Genotyping Strains of Lyme Disease Agents Directly From Ticks, Blood, or Tissue

Barbour, Alan G.; Cook, Vanessa J.

doi:10.1007/978-1-4939-7383-5_1

Cited by 10 publications

(4 citation statements)

References 32 publications

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“…burgdorferi strain diversity within single I. scapularis ticks [41]. Consistent with earlier results based on DNA cloning and DNA-DNA hybridization, the results re-affirmed a largely uniform distribution of a diverse set of B. burgdorferi strains identifiable by ~16 major-group ospC alleles in the highly endemic regions of Lyme disease in the Northeast US [26,29].…”

Section: Lack Of Immune Cross-protection Among B Burgdorferi Strainssupporting

confidence: 87%

Maximum Antigen Diversification in a Lyme Bacterial Population and Evolutionary Strategies to Overcome Pathogen Diversity

Akther

Bezrucenkovas

et al. 2020

Preprint

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Natural populations of microbes and their hosts are engaged in an arms race in which microbes diversify to escape host immunity while hosts evolve novel immunity. This co-evolutionary process, known as the “Red Queen” hypothesis, poses a fundamental challenge to the development of broadly effective vaccines and diagnostics against a diversifying pathogen. Based on surveys of natural allele frequencies and experimental immunization of mice, we show minimal antigenic cross-reactivity among natural variants of the outer surface protein C (OspC), a dominant antigen of a Lyme Disease-causing bacterium (Borrelia burgdorferi). To overcome the challenge of OspC antigenic diversity to clinical development of preventive measures, we implemented a number of evolution-based strategies to broaden OspC immunological cross-reactivity. In particular, the centroid algorithm – a genetic algorithm to minimize sequence differences with natural variants – generated synthetic OspC analogs with the greatest promise as diagnostic and vaccine candidates against diverse Lyme pathogen strains coexisting in the Northeast United States. Mechanistically, we propose a model of runaway maximum antigen di-versification (MAD) mediated by amino-acid variations distributed across hypervariable regions on the OspC molecule. Under the MAD model, evolutionary centroids display high cross-reactivity by occupying the central void in the antigenic space excavated by diversifying natural variants. In contrast to the vaccine design based on concatenated epitopes, the centroid algorithm generates analogs of native antigens and is automated. The MAD model and evolution-inspired antigen designs have broad implications for combating diversifying pathogens driven by pathogen-host coevolution.ImportanceMicrobial pathogens rely on molecular diversity of cell surface antigens to escape host immunity. Vaccines based on one antigen variant often fail to protect the host against pathogens carrying other variants. Here we show evolution-based designs of synthetic antigens that are broadly reactive to all natural variants. The evolutionary analogs of a major surface antigen of a Lyme disease bacterium (Borrelia burgdorferi) showed promise as vaccine candidates against diverse pathogen strains coexisting in the endemic areas of Lyme disease in Northeast United States. Our evolution-based computational design is automated, generates molecular analogs of natural antigens, and opens a novel path to combating fast-evolving microbial pathogens.

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Section: Lack Of Immune Cross-protection Among B Burgdorferi Strainssupporting

confidence: 87%

Maximum Antigen Diversification in a Lyme Bacterial Population and Evolutionary Strategies to Overcome Pathogen Diversity

Akther

Bezrucenkovas

et al. 2020

Preprint

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“…For example, whole-genome sequencing showed that the European B. burgdorferi strain BOL26 obtained its ospC and its flanking genes from a conspecific strain through horizontal gene transfer (32). For population samples elsewhere, therefore, it might be necessary to add a 2nd locus for strain identification (57). One complemental genetic marker could be the rRNA spacer (rrs-rrlA), which is a single-copy and highly variable locus (33).…”

Section: Discussionmentioning

confidence: 99%

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

Wan

Akther

et al. 2018

J Clin Microbiol

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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 (). 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 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 A supporting bioinformatics pipeline has been developed. The newly designed pair of universal 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.

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“…We used the Qiagen DNeasy Blood and Tissue Kit (Qiagen, Hilden, Germany) to extract total nucleic acids from 188 individual flies (S1 Text). First, flies were subjected to a pre-treatment step that has been used widely to extract DNA from ticks [33][34][35][36]. Then, we proceeded with extraction following the manufacturer's protocol.…”

Section: Nucleic Acid Extractionmentioning

confidence: 99%

Sanitation in urban areas may limit the spread of antimicrobial resistance via flies

Capone,

Cumming,

Flemister

et al. 2024

PLoS ONE

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Synanthropic filth flies are common where sanitation is poor and fecal wastes are accessible to them. These flies have been proposed as mechanical vectors for the localized transport of fecal microbes including antimicrobial resistant (AMR) organisms and associated antimicrobial resistance genes (ARGs), increasing exposure risks. We evaluated whether an onsite sanitation intervention in Maputo, Mozambique reduced the concentration of enteric bacteria and the frequency of detection of ARGs carried by flies collected in household compounds of low-income neighborhoods. Additionally, we assessed the phenotypic resistance profile of Enterobacteriaceae isolates recovered from flies during the pre-intervention phase. After fly enumeration at study compounds, quantitative polymerase chain reaction was used to quantify an enteric 16S rRNA gene (i.e., specific to a cluster of phylotypes corresponding to 5% of the human fecal microflora), 28 ARGs, and Kirby Bauer Disk Diffusion of Enterobacteriaceae isolates was utilized to assess resistance to eleven clinically relevant antibiotics. The intervention was associated with a 1.5 log10 reduction (95% confidence interval: -0.73, -2.3) in the concentration of the enteric 16S gene and a 31% reduction (adjusted prevalence ratio = 0.69, [0.52, 0.92]) in the mean number of ARGs per fly compared to a control group with poor sanitation. This protective effect was consistent across the six ARG classes that we detected. Enterobacteriaceae isolates–only from the pre-intervention phase–were resistant to a mean of 3.4 antibiotics out of the eleven assessed. Improving onsite sanitation infrastructure in low-income informal settlements may help reduce fly-mediated transmission of enteric bacteria and the ARGs carried by them.

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Genotyping Strains of Lyme Disease Agents Directly From Ticks, Blood, or Tissue

Cited by 10 publications

References 32 publications

Maximum Antigen Diversification in a Lyme Bacterial Population and Evolutionary Strategies to Overcome Pathogen Diversity

Maximum Antigen Diversification in a Lyme Bacterial Population and Evolutionary Strategies to Overcome Pathogen Diversity

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

Sanitation in urban areas may limit the spread of antimicrobial resistance via flies

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