Development of a PRRSV Detection Assay Using Multienzyme Isothermal Rapid Amplification and Genetic Analysis of PRRSV in Southwest China

IF 3.5 2区农林科学 Q2 INFECTIOUS DISEASES

Transboundary and Emerging Diseases Pub Date : 2025-04-25 DOI:10.1155/tbed/5428900

Juan Zhang, Xinrong Wang, Jun Zhou, Junhai Zhu, Meiyu Jia, Longxiang Zhang, Nan Yan, Lizhi Fu, Yue Wang

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Abstract

Porcine reproductive and respiratory syndrome (PRRS) is a highly contagious viral disease that causes substantial economic losses in the swine industry. This study aimed to develop a PRRS virus (PRRSV) detection assay using multienzyme isothermal rapid amplification (MIRA) and to analyze the genetic variation of PRRSV in Southwest China. A total of 13,863 samples, including blood and lung tissues from pigs suspected of PRRSV infection, were collected. The MIRA assay was designed with primers and probes targeting conserved regions of the PRRSV-M gene, demonstrating high specificity with no cross-reactivity to other swine pathogens and an estimated detection threshold sensitivity of 1.0 copy/μL. Prevalence analysis revealed that, although vaccinated pigs showed relatively high antibody levels, the virus continued to circulate, particularly in unvaccinated herds. Genetic analysis of the predominant PRRSV strains indicated an increasing prevalence of NADC30-like strains and notable genetic variation in genes such as ORF5 and nonstructural protein 2 (NSP2), including amino acid deletions and alterations of glycosylation sites. Recombination events were also observed in some isolates. These findings provide essential insights into the epidemiology and genetic diversity of PRRSV in Southwest China, contributing critical data for the development of more effective control and prevention strategies.

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西南地区PRRSV多酶等温快速扩增检测方法的建立及基因分析

猪繁殖与呼吸综合征（PRRS）是一种传染性极强的病毒性疾病，给养猪业造成了巨大的经济损失。本研究旨在利用多酶等温快速扩增（MIRA）技术开发猪繁殖与呼吸综合征病毒（PRRSV）检测方法，并分析中国西南地区猪繁殖与呼吸综合征病毒的遗传变异。共采集了 13863 份样本，包括疑似感染 PRRSV 的猪的血液和肺组织。MIRA 检测方法采用针对 PRRSV-M 基因保守区的引物和探针设计，特异性高，与其他猪病原体无交叉反应，估计检测阈灵敏度为 1.0 拷贝/μL。流行率分析表明，尽管接种疫苗的猪只表现出相对较高的抗体水平，但病毒仍在继续流行，尤其是在未接种疫苗的猪群中。对主要 PRRSV 株系的遗传分析表明，NADC30 样株系的流行率越来越高，ORF5 和非结构蛋白 2 (NSP2) 等基因的遗传变异显著，包括氨基酸缺失和糖基化位点的改变。在一些分离株中还观察到重组事件。这些发现为了解中国西南地区 PRRSV 的流行病学和遗传多样性提供了重要信息，为制定更有效的防控策略提供了关键数据。

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来源期刊

Transboundary and Emerging Diseases 农林科学-传染病学

CiteScore

8.90

自引率

9.30%

发文量

350

审稿时长

1 months

期刊介绍： Transboundary and Emerging Diseases brings together in one place the latest research on infectious diseases considered to hold the greatest economic threat to animals and humans worldwide. The journal provides a venue for global research on their diagnosis, prevention and management, and for papers on public health, pathogenesis, epidemiology, statistical modeling, diagnostics, biosecurity issues, genomics, vaccine development and rapid communication of new outbreaks. Papers should include timely research approaches using state-of-the-art technologies. The editors encourage papers adopting a science-based approach on socio-economic and environmental factors influencing the management of the bio-security threat posed by these diseases, including risk analysis and disease spread modeling. Preference will be given to communications focusing on novel science-based approaches to controlling transboundary and emerging diseases. The following topics are generally considered out-of-scope, but decisions are made on a case-by-case basis (for example, studies on cryptic wildlife populations, and those on potential species extinctions): Pathogen discovery: a common pathogen newly recognised in a specific country, or a new pathogen or genetic sequence for which there is little context about — or insights regarding — its emergence or spread. Prevalence estimation surveys and risk factor studies based on survey (rather than longitudinal) methodology, except when such studies are unique. Surveys of knowledge, attitudes and practices are within scope. Diagnostic test development if not accompanied by robust sensitivity and specificity estimation from field studies. Studies focused only on laboratory methods in which relevance to disease emergence and spread is not obvious or can not be inferred (“pure research” type studies). Narrative literature reviews which do not generate new knowledge. Systematic and scoping reviews, and meta-analyses are within scope.