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Ascaridia galli  is an important nematode that causes ascaridiasis in free-range and indoor system chicken farms. Infection with  A. galli  may damage the intestinal mucosa and inhibit nutrient absorption, leading to a reduced growth rate, weight loss and a decreased egg production. Consequently,  A. galli  infection is a significant health problem in chickens. In this study, we developed a loop-mediated isothermal amplification coupled with a lateral flow dipstick (LAMP-LFD) assay for the visual detection of  A. galli  eggs in faecal samples. The LAMP-LFD assay consists of six primers and one DNA probe that recognize the internal transcribed spacer 2 (ITS2) region; it can be performed within 70 min and the results can be interpreted with the naked eye. Using the LAMP-LFD assay developed in this study,  A. galli  DNA was specifically amplified without any cross-reactions with other related parasites ( Heterakis gallinarum ,  Raillietina echinobothrida ,  R. tetragona ,  R. cesticillus ,  Cotugnia  sp.,  Echinostoma miyagawai ) and definitive hosts ( Gallus gallus domesticus ,  Anas platyrhynchos domesticus ). The minimum detectable DNA concentration was 5 pg/μl, and the detectable egg count was 50 eggs per reaction. The assay can be performed in a water bath, without the need for  post-mortem  morphological investigations and laboratory instruments. It is therefore a viable alternative for the detection of  A. galli  in chicken faeces and can replace classical methods in field screening for epidemiological investigations, veterinary health and poultry farming management. RESEARCH HIGHLIGHTSThis is the first study using the LAMP-LFD assay for  Ascaridia galli  detection.The results can be observed by the naked eye.The developed assay can be used to detect  Ascaridia galli  eggs in faecal samples. This is the first study using the LAMP-LFD assay for  Ascaridia galli  detection. The results can be observed by the naked eye. The developed assay can be used to detect  Ascaridia galli  eggs in faecal samples.

avian pathology

Feasibility of a DNA biosensor assay based on loop-mediated isothermal amplification combined with a lateral flow dipstick assay for the visual detection of  Ascaridia galli  eggs in faecal samples