The Detection and Differentiation of Pigeon Adenovirus Types 1 and 2 via a High-Resolution Melting Curve Platform
Two main adenoviral diseases have been described in pigeons: pigeon adenovirus type 1 (PiAdV-1) and pigeon adenovirus type 2 (PiAdV-2), which belong to the genus <i>Aviadenovirus</i> under the family <i>Adenoviridae</i>. PiAdV-1 and PiAdV-2 are highly pathogenic to pigeons, l...
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MDPI AG
2025-06-01
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| author | Shuyu Chen Wenyu Zhang Zhiwang Tang Tingting Lu Chunhe Wan Wensong Jin Jiayu Li |
| author_facet | Shuyu Chen Wenyu Zhang Zhiwang Tang Tingting Lu Chunhe Wan Wensong Jin Jiayu Li |
| author_sort | Shuyu Chen |
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| description | Two main adenoviral diseases have been described in pigeons: pigeon adenovirus type 1 (PiAdV-1) and pigeon adenovirus type 2 (PiAdV-2), which belong to the genus <i>Aviadenovirus</i> under the family <i>Adenoviridae</i>. PiAdV-1 and PiAdV-2 are highly pathogenic to pigeons, leading to considerable losses worldwide. To date, there is little information on the epidemiological distribution of PiAdV-1 and PiAdV-2 in pigeons due to the lack of detection and differentiation platforms for these two viruses. High-resolution melting technology (HRM) has been widely used for developing detection and differentiation platforms, with the melting profile based on the GC content in the real-time PCR (qPCR-HRM) system. This study designed and synthesized a pair of specific primers on the basis of the characteristic variations of the 52K genes of PiAdV-1 and PiAdV-2, then the detection and differentiation qPCR-HRM platform was established after conditional optimization. The results showed that this method had good specificity; it could only specifically detect PiAdV-1 and PiAdV-2, with no cross-reaction with other pigeon-origin pathogens that occur in pigeons. This method had high sensitivity, with the lowest detection limits at 57 copies/µL (for PiAdV-1) and 56 copies/µL (for PiAdV-2). This method had good intra-group and inter-group coefficients of variation, both of which were less than 1.5%. Field samples for the epidemiological surveillance and investigation data of PiAdV-1 and PiAdV-2 were checked. We found only PiAdV-2-positive samples in meat pigeons, but the percentages of PiAdV-1-positive, PiAdV-2-positive, and coinfection-positive samples among the racing pigeons were 5.71%, 14.29%, and 2.86%, respectively. To our knowledge, this is the first report for the simultaneous detection and differentiation of PiAdV-1 and PiAdV-2 using the qPCR-HRM platform. Our study also provided evidence of PiAdV-1 and PiAdV-2 coinfection in racing pigeons, but further studies are needed. |
| format | Article |
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| institution | DOAJ |
| issn | 2076-2607 |
| language | English |
| publishDate | 2025-06-01 |
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| series | Microorganisms |
| spelling | doaj-art-ce2cc4fa2da541b9854741a6329bfd9e2025-08-20T03:16:21ZengMDPI AGMicroorganisms2076-26072025-06-01136133110.3390/microorganisms13061331The Detection and Differentiation of Pigeon Adenovirus Types 1 and 2 via a High-Resolution Melting Curve PlatformShuyu Chen0Wenyu Zhang1Zhiwang Tang2Tingting Lu3Chunhe Wan4Wensong Jin5Jiayu Li6School of Life Sciences, Fujian Agriculture and Forestry University, Fuzhou 350002, ChinaSchool of Life Sciences, Fujian Agriculture and Forestry University, Fuzhou 350002, ChinaSchool of Life Sciences, Fujian Agriculture and Forestry University, Fuzhou 350002, ChinaSchool of Life Sciences, Fujian Agriculture and Forestry University, Fuzhou 350002, ChinaInstitute of Animal Husbandry and Veterinary Medicine, Fujian Academy of Agricultural Sciences, Fuzhou 350013, ChinaSchool of Life Sciences, Fujian Agriculture and Forestry University, Fuzhou 350002, ChinaSchool of Life Sciences, Fujian Agriculture and Forestry University, Fuzhou 350002, ChinaTwo main adenoviral diseases have been described in pigeons: pigeon adenovirus type 1 (PiAdV-1) and pigeon adenovirus type 2 (PiAdV-2), which belong to the genus <i>Aviadenovirus</i> under the family <i>Adenoviridae</i>. PiAdV-1 and PiAdV-2 are highly pathogenic to pigeons, leading to considerable losses worldwide. To date, there is little information on the epidemiological distribution of PiAdV-1 and PiAdV-2 in pigeons due to the lack of detection and differentiation platforms for these two viruses. High-resolution melting technology (HRM) has been widely used for developing detection and differentiation platforms, with the melting profile based on the GC content in the real-time PCR (qPCR-HRM) system. This study designed and synthesized a pair of specific primers on the basis of the characteristic variations of the 52K genes of PiAdV-1 and PiAdV-2, then the detection and differentiation qPCR-HRM platform was established after conditional optimization. The results showed that this method had good specificity; it could only specifically detect PiAdV-1 and PiAdV-2, with no cross-reaction with other pigeon-origin pathogens that occur in pigeons. This method had high sensitivity, with the lowest detection limits at 57 copies/µL (for PiAdV-1) and 56 copies/µL (for PiAdV-2). This method had good intra-group and inter-group coefficients of variation, both of which were less than 1.5%. Field samples for the epidemiological surveillance and investigation data of PiAdV-1 and PiAdV-2 were checked. We found only PiAdV-2-positive samples in meat pigeons, but the percentages of PiAdV-1-positive, PiAdV-2-positive, and coinfection-positive samples among the racing pigeons were 5.71%, 14.29%, and 2.86%, respectively. To our knowledge, this is the first report for the simultaneous detection and differentiation of PiAdV-1 and PiAdV-2 using the qPCR-HRM platform. Our study also provided evidence of PiAdV-1 and PiAdV-2 coinfection in racing pigeons, but further studies are needed.https://www.mdpi.com/2076-2607/13/6/1331PiAdV-1PiAdV-2qPCR-HRM platformdetection and differentiationepidemiological investigationcoinfection |
| spellingShingle | Shuyu Chen Wenyu Zhang Zhiwang Tang Tingting Lu Chunhe Wan Wensong Jin Jiayu Li The Detection and Differentiation of Pigeon Adenovirus Types 1 and 2 via a High-Resolution Melting Curve Platform Microorganisms PiAdV-1 PiAdV-2 qPCR-HRM platform detection and differentiation epidemiological investigation coinfection |
| title | The Detection and Differentiation of Pigeon Adenovirus Types 1 and 2 via a High-Resolution Melting Curve Platform |
| title_full | The Detection and Differentiation of Pigeon Adenovirus Types 1 and 2 via a High-Resolution Melting Curve Platform |
| title_fullStr | The Detection and Differentiation of Pigeon Adenovirus Types 1 and 2 via a High-Resolution Melting Curve Platform |
| title_full_unstemmed | The Detection and Differentiation of Pigeon Adenovirus Types 1 and 2 via a High-Resolution Melting Curve Platform |
| title_short | The Detection and Differentiation of Pigeon Adenovirus Types 1 and 2 via a High-Resolution Melting Curve Platform |
| title_sort | detection and differentiation of pigeon adenovirus types 1 and 2 via a high resolution melting curve platform |
| topic | PiAdV-1 PiAdV-2 qPCR-HRM platform detection and differentiation epidemiological investigation coinfection |
| url | https://www.mdpi.com/2076-2607/13/6/1331 |
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