Sex identification of sun conure (Aratinga solstitialis) using loop-mediated isothermal amplification of W and Z spindlin chromosomes

Abstract

Background and Aim: The sun conure (Aratinga solstitialis), a bird belonging to the Psittaciformes family, is a popular pet because of its bright color and beautiful appearance. The sun conure is a monomorphic bird with similar appearances between males and females, making sex identification difficult by observing the external morphology. Therefore, molecular techniques are utilized. Loop-mediated isothermal amplification (LAMP) is a molecular technique that is often applied for sex identification in birds and is a quick and simple method that can be used in the field. This study used the LAMP technique to improve sex identification in sun conures by observing the color change of hydroxy naphthol blue. Materials and Methods: Two primer sets, SunSpin-W and SunSpin-Z, were designed for sex identification in sun conures using the LAMP technique specific to the spindlin gene. The developed LAMP reaction was tested for optimal conditions, sensitivity, and specificity compared with the polymerase chain reaction (PCR) technique. Results: The SunSpin-W primer set amplified only female birds, whereas the SunSpin-Z primer set amplified DNA from both male and female birds. The primer sets were optimized at 62°C for 45 min. A positive result was visible to the naked eye from the color change of the reaction. In the LAMP assay, the lowest detectable concentration was 10 pg/μL and in the PCR assay, it was 1 ng/μL, while a 100% accuracy rate in sex identification was observed when comparing the LAMP assay results with the PCR assay. Conclusion: This study successfully developed a LAMP technique for sex identification of sun conure, which took 45 min to complete and can be expanded for use in the field.

Keywords

• loop-mediated isothermal amplification
• sex identification
• spindlin gene
• sun conure