Development of a Rapid Sex Identification Method for Newborn Pigeons Using Recombinase Polymerase Amplification and a Lateral-Flow Dipstick on Farm

Abstract

According to pigeon racing rules in Taiwan, the pigeon raiser must decide which juveniles will be chosen as soon as possible. Differentiating the sex of young pigeons based on appearances, and other traditional methods, can be time-consuming and require several pieces of equipment. Recombinase polymerase amplification (RPA) combined with a lateral-flow dipstick (LFD) could further simplify the presentation of amplification results. A designed reverse primer and probe were labeled with biotin and FAM (fluorescein), respectively, to serve as ligands in the LFD. With the addition of a designed forward primer, the RPA-LFD can be used to perform sex identification of pigeon DNA. The optimal conditions were determined to require at least 6.3 pg of the DNA template, a temperature of 37 °C, and a reaction time of at least 20 min. Under these conditions, the test band area on the strip appeared as a dark color if the sample contained female template DNA, whereas the male DNA samples did not produce any test signal in any of the conditions. The results of random samples using RPA-LFD under the optimal conditions agreed with the results of the same samples determined by PCR-agarose gel electrophoresis. The approach in this study represents a rapid and accurate method for pigeon sexing.

The development rule of feathers and application of hair root tissue in sex identification of Yangzhou geese

“…At present, this method has achieved early sex identification in chickens ( Ayers et al, 2015 ), ducks ( Li et al, 2015 ), pigeons ( Abou Elez et al, 2023 ), partridges ( Jia et al, 2023 ), ostriches ( Huang C and Huang M, 2008 ), and other species. There are two homologous copies of the CHD gene, of which CHD1-W is linked to the W chromosome and CHD1-Z is linked to the Z chromosome ( Lai et al, 2022 ). The intron sizes of the two homologous gene copies differ considerably.…”

Sex identification methods of birds: a review

“…Recombinase polymerase amplification combined with a lateral-flow dipstick (RPA-LFD) analysis was developed to determine the sex of juvenile pigeon as a less equipped on farm approach and obtained 100 % accuracy in sex determination. It can be practiced rapidly compared with the other PCR-based techniques without diminishing the accuracy of the determination as well (Lai et al, 2022).…”

“…The scenario of determining the sex in animal kingdom exhibits an amazingly higher degree of variability though the accurate sex determination and differentiation are necessary for all sexually reproducing species for their survival (Wilhelm et al, 2007). Birds can be differentiated into two main categories namely dimorphic and monomorphic are based on the general characteristics of male and female birds whereas dimorphic birds could be obviously differentiated into male and female based on their appearance (Lai et al, 2022). There is no distinct sexual dimorphism in approximately 60 % of the bird species and it is hard to distinguish the gender solely by using the morphology (Griffiths, 2000;Wu et al, 2007).…”

“…It would be beneficial while studying the bird populations, behavior, evolution, and wild fauna management. Furthermore, it plays an important role in improving the reproduction planning by analyzing the breeding strategies and additionally provides an added advantages in forensic cases (Lai et al, 2022). Moreover, Chen et al (2022) stated that the sex determination of chicken is very complex and precisely regulated by multiple genes.…”

Fast and Simple Molecular Test for Sex Determination of the Monomorphic Eudromia elegans Individuals

“…This difficulty is not just an inconvenience for researchers; it has significant implications for various aspects of avian biology, including understanding mating systems, population genetics, and conservation strategies. Accurate sex determination within a population allows researchers to investigate patterns of sex ratio, reproductive success, and gene flow, which are crucial for an effective understanding of evolutionary dynamics, conservation management, and added advantages in forensic cases [ 9 , 10 , 11 , 12 , 13 , 14 ].…”