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Laboratory evaluation of molecular xenomonitoring using mosquito and tsetse fly excreta/feces to amplify Plasmodium, Brugia, and Trypanosoma DNA

Pilotte, Nils, Cook, Darren, Pryce, Joseph, Zulch, Michael F., Minetti, Corrado ORCID: https://orcid.org/0000-0001-7862-4874, Reimer, Lisa ORCID: https://orcid.org/0000-0002-9711-4981 and Williams, Steven A. (2020) 'Laboratory evaluation of molecular xenomonitoring using mosquito and tsetse fly excreta/feces to amplify Plasmodium, Brugia, and Trypanosoma DNA'. Gates Open Research, p. 1734.

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Abstract

Background: Results from an increasing number of studies suggest that mosquito excreta/feces (E/F) testing has considerable potential to serve as a supplement for traditional molecular xenomonitoring techniques. However, as the catalogue of possible use-cases for this methodology expands, and the list of amenable pathogens grows, a number of fundamental methods-based questions remain. Answering these questions is critical to maximizing the utility of this approach and to facilitating its successful implementation as an effective tool for molecular xenomonitoring.
Methods: Utilizing E/F produced by mosquitoes or tsetse flies experimentally exposed to Brugia malayi, Plasmodium falciparum, or Trypanosoma brucei brucei, factors such as limits of detection, throughput of testing, adaptability to use with competent and incompetent vector species, and effects of additional blood feedings post parasite-exposure were evaluated. Two platforms for the detection of pathogen signal (quantitative real-time PCR and digital PCR (dPCR)) were also compared, with strengths and weaknesses examined for each.
Results: Experimental results indicated that high throughput testing is possible when evaluating mosquito E/F for the presence of either B. malayi or P. falciparum from both competent and incompetent vector mosquito species. Furthermore, following exposure to pathogen, providing mosquitoes with a second, uninfected bloodmeal did not expand the temporal window for E/F collection during which pathogen detection was possible. However, this collection window did appear longer in E/F collected from tsetse flies following exposure to T. b. brucei. Testing also suggested that dPCR may facilitate detection through its increased sensitivity. Unfortunately, logistical obstacles will likely make the large-scale use of dPCR impractical for this purpose.
Conclusions: By examining many E/F testing variables, expansion of this technology to a field-ready platform has become increasingly feasible. However, translation of this methodology from the lab to the field will first require field-based pilot studies aimed at assessing the efficacy of E/F screening.

Item Type: Article
Subjects: QU Biochemistry > Proteins. Amino Acids. Peptides > QU 58.5 DNA.
QX Parasitology > Protozoa > QX 135 Plasmodia
QX Parasitology > Helminths. Annelida > QX 301 Filarioidea
QX Parasitology > Insects. Other Parasites > QX 510 Mosquitoes
QY Clinical Pathology > QY 25 Laboratory techniques and procedure
WC Communicable Diseases > Tropical and Parasitic Diseases > WC 705 Trypanosomiasis
Faculty: Department: Biological Sciences > Vector Biology Department
Digital Object Identifer (DOI): https://doi.org/10.12688/gatesopenres.13093.2
Depositing User: Stacy Murtagh
Date Deposited: 22 Feb 2021 16:21
Last Modified: 22 Feb 2021 16:21
URI: https://archive.lstmed.ac.uk/id/eprint/17033

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