Exploring the utility of circulating miRNAs as diagnostic biomarkers of fasciolosis

Sumaiya Chowdhury ¹, Alison Ricafrente ¹, Krystyna Cwiklinski ^2,3

⁰The School of Life Sciences, University of Technology, Sydney, Australia.

Scientific Reports +

|

March 29, 2024

View abstract on PubMed

Summary

Early detection of parasitic infections like fasciolosis in sheep is crucial. This study identifies specific microRNAs (miRNAs) in sheep blood that can diagnose Fasciola hepatica infection as early as 7 days, improving livestock health management.

Area Of Science

Veterinary Parasitology
Molecular Diagnostics
Livestock Health

Background

Effective management of parasitic infections in livestock, such as Fasciola hepatica (liver fluke), relies on early detection.
Current diagnostic methods for fasciolosis detect infection around three weeks post-infection, after significant liver damage has occurred.
Investigating novel biomarkers for earlier detection is essential to mitigate economic losses and animal suffering.

Purpose Of The Study

To explore the potential of differentially abundant microRNAs (miRNAs) in sheep serum as early diagnostic biomarkers for Fasciola hepatica infection.
To identify specific host and parasite-derived miRNAs associated with pre-hepatic and hepatic stages of infection.
To validate candidate miRNA biomarkers for early detection of fasciolosis in sheep.

Main Methods

Serum samples from infected and non-infected sheep were analyzed using miRNA sequencing to identify differential expression profiles.
Candidate miRNAs (host and parasite-derived) were selected based on sequencing data.
Quantitative reverse transcription polymerase chain reaction (RT-qPCR) was used to validate the expression levels of selected miRNAs in longitudinal serum samples.
Primer design considerations for accurate RT-qPCR were investigated, particularly for parasite-derived miRNAs and their variants (IsomiRs).

Main Results

Distinct miRNA profiles were observed in sheep sera during pre-hepatic and hepatic Fasciola hepatica infection phases.
Six miRNAs (four sheep, two parasite-derived) were identified as potential biomarkers.
Oar-miR-133-5p and oar-miR-3957-5p were validated as promising biomarkers, detecting infection as early as 7 days post-infection.
Sequence variants (IsomiRs) of parasite miRNA (fhe-miR-124-3p) were prevalent, emphasizing the importance of primer design in diagnostic accuracy.

Conclusions

Specific circulating miRNAs serve as sensitive and early biomarkers for Fasciola hepatica infection in sheep.
The identified host miRNAs, oar-miR-133-5p and oar-miR-3957-5p, enable detection of fasciolosis significantly earlier than traditional methods.
miRNA-based diagnostics hold promise for improved early detection and management of parasitic infections in livestock.
Accurate primer design is critical for the reliable detection of parasite-derived miRNAs and their variants in diagnostic assays.