农业生物技术学报2025，Vol.33Issue(5)：1172-1180,9.DOI:10.3969/j.issn.1674-7968.2025.05.020
番鸭小鹅瘟病毒和番鸭细小病毒三引物双重PCR鉴别检测方法的建立与评价
Establishment and Evaluation of Triple-primer Duplex PCR Method for Differential Detection of Muscovy duck-origin goose parvovirus and Musco-vy duck parvovirus
摘要
Abstract
Muscovy duck-origin goose parvovirus(MDGPV)is a novel waterfowl parvovirus generated by genetic recombination of Classical Goose parvovirus(cGPV)and Muscovy Duck parvovirus(MDPV).It is one of the main pathogens causing severe diarrhea and exudative enteritis in young Muscovy ducks(Cairina moschata),leading to a decline in their immunity and an increase in morbidity and mortality.The study aimed to establish a three-primer PCR method for simultaneous detection and differentiation of MDGPV and MDPV and monitor the current epidemic situation of the 2 viruses.Based on the recombination characteristics of the viral capsid protein(VP1)gene of MDGPV and MDPV published in GenBank,3 differential primers were designed and synthesized using the Oligo 7.0 software.Two primers were combined to form 2 sets of PCR reaction systems for amplification,with the target fragment sizes of 493 bp(MDGPV)and 827 bp(MDPV).Standard positive recombinant plasmids of MDGPV and MDPV were constructed as reference materials for the amplification test.The results showed that this study successfully established a three-primer duplex PCR discrimination method capable of simultaneously detecting MDGPV and MDPV.The three-primer duplex PCR method had good repeatability,high specificity and sensitivity,with the minimum detection limits for MDGPV and MDPV being 99.6 and 96.1 copies/μL of total DNA,respectively.The detection results for 102 clinical samples suspected of being infected with waterfowl Parvovirues were as follows:the positive detection rate of MDGPV was 31.37％(32/102),the positive detection rate of MDPV was 7.84％(8/102),the positive detection rate of mixed infection was 1.96％(2/102),which were consistent with the results of SYBR GreenⅠ-based PCR detection and virus isolation for diagnosing waterfowl Parvovirus infections.In summary,the three-primer PCR diagnostic method established in this study has high specificity,good sensitivity,and is a valuable addition to the detection methods for avian Parvovirus diseases.It is helpful for the clinical diagnosis and epidemiological surveillance of MDGPV and MDPV.关键词
番鸭小鹅瘟病毒(MDGPV)/番鸭细小病毒(MDPV)/双重PCRKey words
Muscovy duck-origin goose parvovirus(MDGPV)/Muscovy duck parvovirus(MDPV)/Duplex PCR分类
畜牧业引用本文复制引用
王劭,刘鸿威,程晓霞,朱小丽,江丹丹,肖世峰,陈少莺,陈仕龙..番鸭小鹅瘟病毒和番鸭细小病毒三引物双重PCR鉴别检测方法的建立与评价[J].农业生物技术学报,2025,33(5):1172-1180,9.基金项目
福建省农业高质量发展超越"5511"协同创新工程项目(XTCXGC2021018) （XTCXGC2021018）
福建省农业科学院对外合作项目(DWHZ2024-05) （DWHZ2024-05）
福建省农业科学院科技创新团队建设项目(CXTD2021034) （CXTD2021034）
福建省属公益类科研院所基本科研专项(2022R1026004 （）
2022R1026006 （）
2023R1024003) （）
国家自然科学基金(32302868) （32302868）
中央引导地方科技发展专项(2022L3019) （2022L3019）
