# Application of parallel PCR and DNA sequencing for detecting remnant host DNA from the prior blood meal of Ixodes scapularis

> **NIH NIH R03** · NORTHERN VERMONT UNIVERSITY-JOHNSON · 2021 · $57,705

## Abstract

PROJECT SUMMARY/ABSTRACT
Knowledge of the host species that maintain populations of Ixodes scapularis, the Lyme disease vector,
and Borrelia burgdorferi, the Lyme disease pathogen, is essential to improving preventative strategies for
this debilitating disease that affects an estimated 300,000 people annually in the United States. However,
the majority of research on host species is focused on only two host species - the white-tailed deer
(Odocoileus virginianus) and the white-footed mouse (Peromyscus leucopus), despite documentation of
19 mammals and 60 bird species that are permissive hosts for I. scapularis in the eastern United States.
The specific aim of this study is to test a new amplicon sequencing protocol for determining the identity of
the larval stage blood meal of questing Ixodes scapularis nymphs. We will focus on nymphs because this
is the life stage responsible for most Lyme disease cases in the eastern United States.
This new host detection protocol utilizes parallel PCR with primer pairs targeting 18S, 12S and 16S rDNA
to amplify relatively short DNA gene regions (115 - 294 nt) remnant from the larval stage blood meal, as
well as additional primer sets for detection of common human pathogens (B. burgdorferi, Anaplasma
phagocytophilum and Babesia microti, agents of Lyme disease, Anaplasmosis and human Babesiosis,
respectively). This will be followed by high-throughput sequencing of the amplicons, and pathogen and
taxonomic classification. The protocol will be tested on a unique library of positive controls - DNA isolates
from I. scapularis nymphs for which the larval stage host is known. Additionally, the protocol will be tested
on hundreds of I. scapularis DNA isolates collected in southern Vermont - a region with one of the highest
rates of Lyme disease incidence in the United States - that were collected across nine long-term study
sites from 2015 - 2017. These data will provide needed insights into I. scapularis host use as well as
reservoir capacity, or the absolute contribution of a reservoir host to the prevalence of pathogen infection
in a tick population. This information can be used to prevent Lyme disease transmission by improving
estimates of risk over time and space through improved knowledge of wildlife host contributions to
pathogen transmission, and also by informing management strategies based on mitigating the
contributions of key host species.

## Key facts

- **NIH application ID:** 10407690
- **Project number:** 7R03AI159287-02
- **Recipient organization:** NORTHERN VERMONT UNIVERSITY-JOHNSON
- **Principal Investigator:** William Landesman
- **Activity code:** R03 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2021
- **Award amount:** $57,705
- **Award type:** 7
- **Project period:** 2021-03-08 → 2023-02-28

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10407690

## Citation

> US National Institutes of Health, RePORTER application 10407690, Application of parallel PCR and DNA sequencing for detecting remnant host DNA from the prior blood meal of Ixodes scapularis (7R03AI159287-02). Retrieved via AI Analytics 2026-05-23 from https://api.ai-analytics.org/grant/nih/10407690. Licensed CC0.

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