Establishment of an LNA-TaqMan fluorescent quantitative PCR assay for differential diagnosis of virulent and attenuated strains of short beak and dwarfism syndrome virus.

IF 2.7 2区农林科学 Q1 VETERINARY SCIENCES

Avian Pathology Pub Date : 2026-07-13 DOI:10.1080/03079457.2026.2692641

Su Lin, Xiaoxia Cheng, Shaoying Chen, Shilong Chen, Shao Wang

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Abstract

To enable differential detection of virulent and attenuated strains of duck short beak and dwarfism syndrome virus (SBDSV), we developed a fluorescent quantitative PCR (qPCR) assay based on locked nucleic acid (LNA)-TaqMan probes. This assay utilized a pair of universal primers and two strain-specific LNA-TaqMan probes (labelled with FAM and VIC, respectively) targeting the VP1 gene of SBDSV. Validation results showed that the assay specifically detected SBDSV virulent and attenuated strains without cross-reacting with other common waterfowl viruses. It exhibited high sensitivity, with a minimum detection limit of 6.0 × 10⁰ copies/μl, and excellent reproducibility, with intra- and inter-assay coefficients of variation (CVs) all < 3%. Additionally, viral titre estimates from this qPCR assay showed no significant difference from those obtained via the gold-standard 50% tissue culture infectious dose (TCID₅₀) assay (P > 0.05). In conclusion, this LNA-TaqMan qPCR assay provides a specific, sensitive, and reliable tool for the differential diagnosis of SBDSV virulent and attenuated strains in clinical practice, serving as strong technical support for the prevention, control, and eradication of SBDSV.

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短喙矮化综合征病毒弱毒株与毒株鉴别诊断的LNA-TaqMan荧光定量PCR方法的建立

摘要为了区分鸭短喙矮化综合征病毒（SBDSV）强毒株和弱毒株，建立了一种基于锁定核酸(LNA)-TaqMan探针的荧光定量PCR （qPCR）检测方法。本实验利用一对通用引物和两个菌株特异性lnna - taqman探针（分别标记为FAM和VIC）靶向SBDSV的VP1基因。验证结果表明，该方法能特异性检测出SBDSV毒株和弱毒株，与其他常见水禽病毒无交叉反应。该方法灵敏度高，最低检出限为6.0 × 10⁰copies/μL，重现性好，测定内和测定间变异系数（CVs）均< 3%。此外，该qPCR检测的病毒滴度估计值与通过金标准50%组织培养感染剂量（TCID₅0）检测（P > 0.05）获得的病毒滴度估计值没有显着差异。综上所述，LNA-TaqMan qPCR检测在临床实践中为SBDSV强毒株和弱毒株的鉴别诊断提供了特异、灵敏、可靠的工具，为SBDSV的预防、控制和根除提供了强有力的技术支持。

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来源期刊

Avian Pathology 农林科学-兽医学

CiteScore

4.50

自引率

10.70%

发文量

审稿时长

1 months

期刊介绍： Avian Pathology is the official journal of the World Veterinary Poultry Association and, since its first publication in 1972, has been a leading international journal for poultry disease scientists. It publishes material relevant to the entire field of infectious and non-infectious diseases of poultry and other birds. Accepted manuscripts will contribute novel data of interest to an international readership and will add significantly to knowledge and understanding of diseases, old or new. Subject areas include pathology, diagnosis, detection and characterisation of pathogens, infections of possible zoonotic importance, epidemiology, innate and immune responses, vaccines, gene sequences, genetics in relation to disease and physiological and biochemical changes in response to disease. First and subsequent reports of well-recognized diseases within a country are not acceptable unless they also include substantial new information about the disease or pathogen. Manuscripts on wild or pet birds should describe disease or pathogens in a significant number of birds, recognizing/suggesting serious potential impact on that species or that the disease or pathogen is of demonstrable relevance to poultry. Manuscripts on food-borne microorganisms acquired during or after processing, and those that catalogue the occurrence or properties of microorganisms, are unlikely to be considered for publication in the absence of data linking them to avian disease.