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...

Full description

Saved in:
Bibliographic Details
Main Authors: Shuyu Chen, Wenyu Zhang, Zhiwang Tang, Tingting Lu, Chunhe Wan, Wensong Jin, Jiayu Li
Format: Article
Language:English
Published: MDPI AG 2025-06-01
Series:Microorganisms
Subjects:
Online Access:https://www.mdpi.com/2076-2607/13/6/1331
Tags: Add Tag
No Tags, Be the first to tag this record!
_version_ 1849705933060440064
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
collection DOAJ
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
id doaj-art-ce2cc4fa2da541b9854741a6329bfd9e
institution DOAJ
issn 2076-2607
language English
publishDate 2025-06-01
publisher MDPI AG
record_format Article
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
work_keys_str_mv AT shuyuchen thedetectionanddifferentiationofpigeonadenovirustypes1and2viaahighresolutionmeltingcurveplatform
AT wenyuzhang thedetectionanddifferentiationofpigeonadenovirustypes1and2viaahighresolutionmeltingcurveplatform
AT zhiwangtang thedetectionanddifferentiationofpigeonadenovirustypes1and2viaahighresolutionmeltingcurveplatform
AT tingtinglu thedetectionanddifferentiationofpigeonadenovirustypes1and2viaahighresolutionmeltingcurveplatform
AT chunhewan thedetectionanddifferentiationofpigeonadenovirustypes1and2viaahighresolutionmeltingcurveplatform
AT wensongjin thedetectionanddifferentiationofpigeonadenovirustypes1and2viaahighresolutionmeltingcurveplatform
AT jiayuli thedetectionanddifferentiationofpigeonadenovirustypes1and2viaahighresolutionmeltingcurveplatform
AT shuyuchen detectionanddifferentiationofpigeonadenovirustypes1and2viaahighresolutionmeltingcurveplatform
AT wenyuzhang detectionanddifferentiationofpigeonadenovirustypes1and2viaahighresolutionmeltingcurveplatform
AT zhiwangtang detectionanddifferentiationofpigeonadenovirustypes1and2viaahighresolutionmeltingcurveplatform
AT tingtinglu detectionanddifferentiationofpigeonadenovirustypes1and2viaahighresolutionmeltingcurveplatform
AT chunhewan detectionanddifferentiationofpigeonadenovirustypes1and2viaahighresolutionmeltingcurveplatform
AT wensongjin detectionanddifferentiationofpigeonadenovirustypes1and2viaahighresolutionmeltingcurveplatform
AT jiayuli detectionanddifferentiationofpigeonadenovirustypes1and2viaahighresolutionmeltingcurveplatform