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

Vet World Vol.17 September-2024 Article - 6

Research Article

Veterinary World, 17(9): 2000-2007

https://doi.org/10.14202/vetworld.2024.2000-2007

Parichart Wancham¹, Sakuna Phatthanakunanan², Siriluk Jala², Kanyakodchanan Woramahatthanon², Supaphen Sripiboon³, and Preeda Lertwatcharasarakul⁴

1. Animal Health and Biomedical Sciences Study Program, Faculty of Veterinary Medicine, Kasetsart University, Bangkok 10900, Thailand.
2. Kamphaeng Saen Veterinary Diagnostic Center, Faculty of Veterinary Medicine, Kasetsart University, Nakorn Pathom 73140, Thailand.
3. Department of Large Animal and Wildlife Clinical Science, Kasetsart University, Nakorn Pathom 73140, Thailand.
4. Department of Pathology, Kasetsart University, Nakorn Pathom 73140, Thailand.

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.

How to cite this article: Wancham P, Phatthanakunanan S, Jala S, Woramahatthanon K, Sripiboon S, and Lertwatcharasarakul P (2024) Sex identification of sun conure (Aratinga solstitialis) using loop-mediated isothermal amplification of W and Z spindlin chromosomes, Veterinary World, 17(9): 2000-2007.

Received: 2024-04-23 Accepted: 2024-08-02 Published online: 2024-09-08

Corresponding author: Preeda Lertwatcharasarakul E-mail: preeda.le@ku.th

DOI: 10.14202/vetworld.2024.2000-2007

Copyright: Wancham, et al. This article is an open access article distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/ by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http:// creativecommons.org/ publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.