Sessions

The aim of this study was to present the complete development process of a reliable LAMP-based method for sex determination in domestic pigeons. Molecular genetic approaches are of particular importance, as morphological traits often do not allow accurate identification of sex in birds. In the first phase, PCR and LAMP primers published by Chan et al. (2012) were tested, but these proved non-functional. Even after re-evaluation and correction against the Columba livia domestica reference genome, satisfactory results could not be achieved, which necessitated the design of ten novel primer sets. Two of these produced interpretable results, and combining multiple sets further improved efficiency, although the required specificity has not yet been reached. Application of the WarmStart® Colorimetric LAMP 2X Master Mix supplemented with betaine identified the ALT_V2 primer set as the most effective at 65 °C with a 50-minute incubation, but further primer refinement and optimization of reaction conditions remain necessary.

In the comparison of three swab-based sampling techniques, no significant differences were found in DNA yield; however, practical limitations were observed: removal of cotton from ear swabs was cumbersome, while cattle-designed swabs proved oversized. Evaluation of the market environment for molecular sex determination revealed that international PCR-based services typically cost HUF 4,000-11,000 per sample (gross), whereas the in-house LAMP procedure can realistically be offered at HUF 2,390-2,950 per sample, with further reductions possible at higher batch sizes. To illustrate practical applicability, a “one-page” farm-level SOP protocol was also developed, serving as a template for field implementation of the finalized method after primer optimization, allowing sample collection, reaction setup, and evaluation outside conventional laboratory conditions.

In conclusion, combining LAMP technology with noninvasive sampling represents a promising approach for sex determination in pigeons. Although the current system does not yet reach the specificity required for routine application, the demonstrated economic advantages and the practical feasibility supported by the farm protocol highlight the need for further development, and confirm that LAMP could become a fast, cost-effective, and reliable alternative for pigeon breeders in determining the sex of breeding candidates.