Maximizing ddPCR Efficiency for Precise Roundworm Surveillance in Laying Hens

Background

McMaster and flotation techniques are widely employed in veterinary parasitology as diagnostic tools to detect and quantify gastrointestinal parasites. These techniques are pivotal in detecting and assessing the infection level within an individual or a group of animals, aiding in the formulation of effective treatment and control strategies. However, while these methods are valuable, they may have limitations such as variations in sensitivity based on the type of parasites present and the technique's ability to detect certain parasite stages. As such, complementary diagnostic methods or repeated sampling might be necessary.

Digital droplet polymerase chain reaction (ddPCR) presents several advantages over traditional McMaster and flotation techniques in the field of veterinary parasitology, particularly in the context of detecting and quantifying parasites. It allows for the absolute quantification of target nucleic acids, enabling the detection of very low parasite loads that might be missed by conventional methods. Unlike McMaster and flotation techniques that provide estimations in terms of eggs per gram (EPG) of feces, ddPCR offers absolute quantification of the target genetic material. ddPCR allows for multiplexing, enabling the simultaneous detection and quantification of multiple parasite species or targets within a single reaction. This feature can be particularly advantageous in mixed infections or when monitoring various parasites concurrently.

Methods

Fecal samples from contaminated farms will be first analyzed by flotation and McMaster methods. The same samples will be analyzed by ddPCR. Samples for ddPCR will be prepared according to different protocols. In the end results of the three methods will be compared and possible correlation and agreement between the three methods will be assessed.   

Time frame

Starting September 2024. Suitable for 30hp and 45 hp project.