中国兽医科学2025，Vol.55Issue(12)：1635-1642,8.DOI:10.16656/j.issn.1673-4696.2025.0219
类NADC30-like株和HP-PRRSV鉴别诊断RT-PCR方法的建立及应用
Establishment and application of RT-PCR method for the differential diagnosis of NADC30-like strains and HP-PRRSV
摘要
Abstract
This study aims to establish an RT-PCR method for the differential diagnosis of NADC30-like porcine reproductive and respiratory syndrome virus and highly pathogenic PRRSV(HP-PRRSV),pro-viding a basis for the detection,prevention and control of this disease.Based on the nucleotide se-quence characteristics of the PRRSV Nsp2 gene,an RT-PCR method capable of simultaneously identifying HP-PRRSV and NADC30-like strains was designed.It was found that the sizes of the Nsp2 gene fragments am-plified by this method in the HP-PRRSV and NADC30-like strains were 931 bp and 628 bp respectively,and the minimum detection limits were 472 copies/μL and 132 copies/μL respectively.No specific bands were amplified for other gene subtypes of PRRSV and pathogen.For this method,the dominant strains were iso-lated and purified.A total of 196 clinical samples from pig farms in some areas of Gansu Province were tested.It was found that 28 samples were positive for HP-PRRSV nucleic acid,15 samples were positive for NADC30-like nucleic acid,and 3 samples were positive for HP-PRRSV+NADC30-like nucleic acid.RT-PCR detection of the ORF5 gene revealed that the number of positive samples for HP-PRRSV and NADC30-like nucleic acids was 37 and 9 respectively,but no mixed infection samples were found.The domi-nant strain was isolated and the virus with stable passage was obtained.After purification,virus parti-cles with high purity and intact structure were obtained.The RT-PCR method established in this study can be used for the rapid differential diagnosis and molecular epidemiological investigation of HP-PRRSV and NADC30-like strains.However,it exhibited low sensitivity and which can not detect other PRRSV sub-type strains.关键词
HP-PRRSV/NADC30-like/鉴别诊断/RT-PCR/分离纯化Key words
HP-PRRSV/NADC30-like/differential diagnosis/RT-PCR/isolation and purification分类
农业科技引用本文复制引用
乔璐,柳海云,杨健,黄国梁,郭慧琛,孙世琪,牛家强..类NADC30-like株和HP-PRRSV鉴别诊断RT-PCR方法的建立及应用[J].中国兽医科学,2025,55(12):1635-1642,8.基金项目
兰州市人才项目(2024-QN-3,2023-RC-3) （2024-QN-3,2023-RC-3）
甘肃省自然科学基金项目(23JRRA551,24JRRA012) （23JRRA551,24JRRA012）
甘肃省青年科技基金项目(25JRRA442) （25JRRA442）
