Using Recombinase-Aid Amplification Combined with <i>Pyrococcus furiosus</i> Argonaute for Rapid Sex Identification in Flamingo (<i>Phoenicopteridae</i>)
Shenluan Tan,
Fanwen Zeng,
Wanhuan Zhong,
Tanzipeng Chen,
Xuanjiao Chen,
Li Li,
Hengxi Wei,
Shouquan Zhang
Affiliations
Shenluan Tan
State Key Laboratory of Swine and Poultry Breeding Industry, National Engineering Research Center for Breeding Swine Industry, Guangdong Provincial Key Laboratory of Agro-Animal Genomics and Molecular Breeding, College of Animal Science, South China Agricultural University, Guangzhou 510642, China
Fanwen Zeng
Guangzhou Zoo & Guangzhou Wildlife Research Center, Guangzhou 510075, China
Wanhuan Zhong
Kunming Institute of Zoology, Chinese Academy of Sciences, Kunming 650201, China
Tanzipeng Chen
Guangzhou Zoo & Guangzhou Wildlife Research Center, Guangzhou 510075, China
Xuanjiao Chen
Guangzhou Zoo & Guangzhou Wildlife Research Center, Guangzhou 510075, China
Li Li
State Key Laboratory of Swine and Poultry Breeding Industry, National Engineering Research Center for Breeding Swine Industry, Guangdong Provincial Key Laboratory of Agro-Animal Genomics and Molecular Breeding, College of Animal Science, South China Agricultural University, Guangzhou 510642, China
Hengxi Wei
State Key Laboratory of Swine and Poultry Breeding Industry, National Engineering Research Center for Breeding Swine Industry, Guangdong Provincial Key Laboratory of Agro-Animal Genomics and Molecular Breeding, College of Animal Science, South China Agricultural University, Guangzhou 510642, China
Shouquan Zhang
State Key Laboratory of Swine and Poultry Breeding Industry, National Engineering Research Center for Breeding Swine Industry, Guangdong Provincial Key Laboratory of Agro-Animal Genomics and Molecular Breeding, College of Animal Science, South China Agricultural University, Guangzhou 510642, China
Flamingos (Phoenicopteridae) are among the oldest birds worldwide and are loved by people for their bright red feathers. In addition, flamingos are sexually monomorphic birds, and distinguishing between males and females is difficult. The polymerase chain reaction (PCR) is widely used for sex identification. However, the PCR method requires a precise thermal cycler in the laboratory and is time-consuming. Therefore, developing a rapid, sensitive, and accurate method to identify the sex of flamingos is crucial. In this study, we established a sex identification system using a recombinase-aided amplification-Pyrococcus furiosus Argonaute (RAA-PfAgo) technique for greater flamingo (Phoenicopterus roseus). The greater flamingo-RAA-PfAgo system can identify unknown-sex greater flamingos in less than 1 h and can be visualized using a fluorescent detector or blue light. The results showed that optimal RAA-PfAgo conditions could detect 0.6 ng of genomic DNA and effectively differentiate between males and females. Random sample evaluations revealed that the system had a 100% coincidence rate compared with conventional PCR. In conclusion, this study provides a sensitive, specific, and accurate reference method for greater flamingo sexing.
