Using Recombinase-Aid Amplification Combined with <i>Pyrococcus furiosus</i> Argonaute for Rapid Sex Identification in Flamingo (<i>Phoenicopteridae</i>)
Flamingos (<i>Phoenicopteridae</i>) 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...
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2024-12-01
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| author | Shenluan Tan Fanwen Zeng Wanhuan Zhong Tanzipeng Chen Xuanjiao Chen Li Li Hengxi Wei Shouquan Zhang |
| author_facet | Shenluan Tan Fanwen Zeng Wanhuan Zhong Tanzipeng Chen Xuanjiao Chen Li Li Hengxi Wei Shouquan Zhang |
| author_sort | Shenluan Tan |
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| description | Flamingos (<i>Phoenicopteridae</i>) 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-<i>Pyrococcus furiosus</i> Argonaute (RAA-<i>Pf</i>Ago) technique for greater flamingo (<i>Phoenicopterus roseus</i>). The greater flamingo-RAA-<i>Pf</i>Ago 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-<i>Pf</i>Ago 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. |
| format | Article |
| id | doaj-art-5d9340006f624da4a481d1bc7d0590d3 |
| institution | Kabale University |
| issn | 2076-2615 |
| language | English |
| publishDate | 2024-12-01 |
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| spelling | doaj-art-5d9340006f624da4a481d1bc7d0590d32025-01-10T13:13:46ZengMDPI AGAnimals2076-26152024-12-01151710.3390/ani15010007Using Recombinase-Aid Amplification Combined with <i>Pyrococcus furiosus</i> Argonaute for Rapid Sex Identification in Flamingo (<i>Phoenicopteridae</i>)Shenluan Tan0Fanwen Zeng1Wanhuan Zhong2Tanzipeng Chen3Xuanjiao Chen4Li Li5Hengxi Wei6Shouquan Zhang7State 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, ChinaGuangzhou Zoo & Guangzhou Wildlife Research Center, Guangzhou 510075, ChinaKunming Institute of Zoology, Chinese Academy of Sciences, Kunming 650201, ChinaGuangzhou Zoo & Guangzhou Wildlife Research Center, Guangzhou 510075, ChinaGuangzhou Zoo & Guangzhou Wildlife Research Center, Guangzhou 510075, ChinaState 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, ChinaState 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, ChinaState 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, ChinaFlamingos (<i>Phoenicopteridae</i>) 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-<i>Pyrococcus furiosus</i> Argonaute (RAA-<i>Pf</i>Ago) technique for greater flamingo (<i>Phoenicopterus roseus</i>). The greater flamingo-RAA-<i>Pf</i>Ago 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-<i>Pf</i>Ago 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.https://www.mdpi.com/2076-2615/15/1/7flamingosex identificationrecombinase-aided amplification (RAA)<i>Pyrococcus furiosus</i> Argonaute (<i>Pf</i>Ago) |
| spellingShingle | Shenluan Tan Fanwen Zeng Wanhuan Zhong Tanzipeng Chen Xuanjiao Chen Li Li Hengxi Wei Shouquan Zhang Using Recombinase-Aid Amplification Combined with <i>Pyrococcus furiosus</i> Argonaute for Rapid Sex Identification in Flamingo (<i>Phoenicopteridae</i>) Animals flamingo sex identification recombinase-aided amplification (RAA) <i>Pyrococcus furiosus</i> Argonaute (<i>Pf</i>Ago) |
| title | Using Recombinase-Aid Amplification Combined with <i>Pyrococcus furiosus</i> Argonaute for Rapid Sex Identification in Flamingo (<i>Phoenicopteridae</i>) |
| title_full | Using Recombinase-Aid Amplification Combined with <i>Pyrococcus furiosus</i> Argonaute for Rapid Sex Identification in Flamingo (<i>Phoenicopteridae</i>) |
| title_fullStr | Using Recombinase-Aid Amplification Combined with <i>Pyrococcus furiosus</i> Argonaute for Rapid Sex Identification in Flamingo (<i>Phoenicopteridae</i>) |
| title_full_unstemmed | Using Recombinase-Aid Amplification Combined with <i>Pyrococcus furiosus</i> Argonaute for Rapid Sex Identification in Flamingo (<i>Phoenicopteridae</i>) |
| title_short | Using Recombinase-Aid Amplification Combined with <i>Pyrococcus furiosus</i> Argonaute for Rapid Sex Identification in Flamingo (<i>Phoenicopteridae</i>) |
| title_sort | using recombinase aid amplification combined with i pyrococcus furiosus i argonaute for rapid sex identification in flamingo i phoenicopteridae i |
| topic | flamingo sex identification recombinase-aided amplification (RAA) <i>Pyrococcus furiosus</i> Argonaute (<i>Pf</i>Ago) |
| url | https://www.mdpi.com/2076-2615/15/1/7 |
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