Application of Loop-Mediated Isothermal Amplification (LAMP) in Sex Identification of Parrots Bred in Egypt

SIMPLE SUMMARY: Loop-mediated isothermal amplification is an enzymatic reaction that allows for specific amplification of a chosen DNA fragment. It works at a constant temperature without the need for expensive equipment. The reaction result can be verified without electrophoresis making it ideal for field application or poorly resourced laboratories. In this work, the authors have compared the LAMP and PCR performance in molecular sex identification of parrots using the DNA extracted from the feathers. The authors examined samples from 21 parrot species. They show that PSI-W primers and LAMP are suitable for sexing 17 species not tested previously. ABSTRACT: Over 400 of the 3800 tropical avian species are endangered or threatened. One of many solutions to conserve animal biodiversity is breeding animals in zoos or private animal farms. Animal breeding programs are difficult to implement in species with sexual monomorphism, such as parrots. Molecular biology methods offer a solution to determine the sex of these species. Therefore, in this study, we aimed to test the performance of PCR and LAMP techniques on sex identification for 21 parrot species belonging to three families, i.e., Psittacidae, Cacatuidae, and Psittaculidae. We established a protocol for DNA isolation from feathers in our laboratory and found optimal conditions for PCR and LAMP. We showed that the LAMP method with the use of the PSI-W primers set, developed by Centeno-Cuadros, functions in 17 previously untested species. Moreover, we found that further improvements are required in universal LAMP primers for the detection of parrot DNA, which are necessary for confirmation of the male sex. The LAMP method also proved to be more sensitive for female sex identification in contrast to the reference PCR test. Therefore, we conclude that LAMP is a suitable method for the routine diagnostic sex identification of parrots.