@misc{10481/80923,
year = {2023},
month = {2},
url = {https://hdl.handle.net/10481/80923},
abstract = {Background Trypanosomatid parasites are widely distributed in nature and can have a monoxenous or dixenous
life-cycle. These parasites thrive in a wide number of insect orders, some of which have an important economic and
environmental value, such as bees. The objective of this study was to develop a robust and sensitive real-time quantitative
PCR (qPCR) assay for detecting trypanosomatid parasites in any type of parasitized insect sample.
Methods A TaqMan qPCR assay based on a trypanosomatid-conserved region of the α-tubulin gene was standardized
and evaluated. The limits of detection, sensitivity and versatility of the α-tubulin TaqMan assay were tested and
validated using field samples of honeybee workers, wild bees, bumblebees and grasshoppers, as well as in the human
infective trypanosomatid Leishmania major.
Results The assay showed a detection limit of 1 parasite equivalent/μl and successfully detected trypanosomatids
in 10 different hosts belonging to the insect orders Hymenoptera and Orthoptera. The methodology was also tested
using honeybee samples from four apiaries (n = 224 worker honeybees) located in the Alpujarra region (Granada,
Spain). Trypanosomatids were detected in 2.7% of the honeybees, with an intra-colony prevalence of 0% to 13%. Parasite
loads in the four different classes of insects ranged from 40.6 up to 1.1 × 108
cell equivalents per host.
Conclusions These results show that the α-tubulin TaqMan qPCR assay described here is a versatile diagnostic tool
for the accurate detection and quantification of trypanosomatids in a wide range of environmental settings.},
organization = {Spanish Programme for Knowledge Generation and Scientific and Technological Strengthening of the R+D+I},
organization = {System: Generacion del Conocimiento 2018	PGC2018-098929-A-I00
PID2021-126938OB-I00},
publisher = {BMC},
keywords = {Lotmaria},
keywords = {Crithidia},
keywords = {Leishmania},
keywords = {Prevalence},
keywords = {Epidemiology},
keywords = {Honeybee},
keywords = {Diagnostic},
title = {Development of a TaqMan qPCR assay for trypanosomatid multi‑species detection and quantification in insects},
doi = {10.1186/s13071-023-05687-3},
author = {Barranco Gómez, Olga and Carreira de Paula, Jessica and Solano Parada, Jennifer and Gómez Moracho, Tamara and Vic Marfil, Ana and Zafra, María and Orantes Bermejo, Francisco José and Osuna Carrillo De Albornoz, Antonio and Pablos Torró, Luis Miguel de},
}