The PERL toolkit: Using sand flies to identify leishmaniasis reservoirs
- 1Vector Molecular Biology Section, Laboratory of Malaria and Vector Research, National Institute of Allergy and Infectious Diseases, National Institutes of Healt
Abstract
Animal reservoirs remain unknown in emerging and most endemic leishmaniasis foci hindering control efforts. Here, we developed a field-applicable PERL (Phlebotomines Establish Reservoirs of Leishmania) toolkit using individual blood fed sand flies (IBF) to identify leishmaniasis reservoirs. Using IBF, we optimized DNA and RNA co-extraction and parasite detection of [≥]1 parasite/s by kDNA qPCR and ssu rRNA RT-qPCR. We then screened IBF for expression of two parasite genes, sherp and HPB, identified by RNAseq as having low-to-absent expression in IBF given a first Leishmania donovani-infected blood meal (IBF-iBM1) and high expression in specimens provided subsequent uninfected blood meals (IBF-BMS+). Linear discriminant analysis of target gene expression classified iBM1 parasites with a predictive accuracy of ~87% and ~82% in membrane- or naturally-fed on hamsters sand flies, respectively. By determining the blood source in specimens determined as IBF-iBM1, the PERL toolkit provides an innovative and practical approach to identification of leishmaniasis reservoirs.
Related
- MDigest: A comprehensive toolkit for detection of allosteric communication from molecular dynamics simulations of biochemical systems
- Tangermeme: a toolkit for understanding cis-regulatory logic using deep learning models
- MeXpose 2.0: A Comprehensive Toolkit for Single-Cell Spatial (Metall-)omicsCovering All Steps from Image Reconstruction to Integrative Analysis.
- CRISPR-HAWK: haplotype- and variant-aware guide design toolkit for CRISPR-Cas.
- CellBouncer, a unified toolkit for single-cell demultiplexing and ambient RNA analysis, reveals hominid mitochondrial incompatibilities
BibTeX
@misc{iniguez2026perl,
title = {{The PERL toolkit: Using sand flies to identify leishmaniasis reservoirs}},
author = {E. Iniguez and P. Huffcutt and T. D. Serafim and P. Cecilio and S. Doh and A. Pugh and J. Doehl and C. Meneses and B. Lambert and J. G. Valenzuela and S. Kamhawi},
howpublished = {bioRxiv},
year = {2026},
month = jun,
doi = {10.64898/2026.06.27.734966},
url = {https://www.biorxiv.org/content/10.64898/2026.06.27.734966v2},
note = {CC0},
}
RIS
TY - UNPB
TI - The PERL toolkit: Using sand flies to identify leishmaniasis reservoirs
AU - E. Iniguez
AU - P. Huffcutt
AU - T. D. Serafim
AU - P. Cecilio
AU - S. Doh
AU - A. Pugh
AU - J. Doehl
AU - C. Meneses
AU - B. Lambert
AU - J. G. Valenzuela
AU - S. Kamhawi
PY - 2026
DA - 2026/06/27
PB - bioRxiv
DO - 10.64898/2026.06.27.734966
UR - https://www.biorxiv.org/content/10.64898/2026.06.27.734966v2
AB - Animal reservoirs remain unknown in emerging and most endemic leishmaniasis foci hindering control efforts. Here, we developed a field-applicable PERL (Phlebotomines Establish Reservoirs of Leishmania) toolkit using individual blood fed sand flies (IBF) to identify leishmaniasis reservoirs. Using IBF, we optimized DNA and RNA co-extraction and parasite detection of [≥]1 parasite/s by kDNA qPCR and ssu rRNA RT-qPCR. We then screened IBF for expression of two parasite genes, sherp and HPB, identified by RNAseq as having low-to-absent expression in IBF given a first Leishmania donovani-infected blood meal (IBF-iBM1) and high expression in specimens provided subsequent uninfected blood meals (IBF-BMS+). Linear discriminant analysis of target gene expression classified iBM1 parasites with a predictive accuracy of ~87% and ~82% in membrane- or naturally-fed on hamsters sand flies, respectively. By determining the blood source in specimens determined as IBF-iBM1, the PERL toolkit provides an innovative and practical approach to identification of leishmaniasis reservoirs.
ER -