江西农业学报2024，Vol.36Issue(3)：61-66,6.DOI:10.19386/j.cnki.jxnyxb.2024.03.010
H1亚型禽流感病毒RT-RPA-LFD快速检测方法的建立及应用
Development of Rapid Detection Method for H1 Subtype Avian Influenza Virus RT-RPA-LFD
摘要
Abstract
Taking the HA gene of H1 subtype AIV as the research object,recombinant polymerase nucleic acid isothermal amplification technology(RPA)was used to select a set of primers by RT-PCR method,and the temperature of the reaction system and the probe and primer concentration were optimized.The sensitivity and specificity of the method were tested using the optimized reaction system,and the optimized system was used to explore the shortest reaction time.Based on the results of lateral flow dipstick(LFD),the RT-RPA-LFD rapid detection method for H1 subtype AIV was established.The results showed that the rapid RT-RPA-LFD assay for detection of H1 subtype AIV RNA could detect 1.5×102 copies/response of H1 subtype AIV RNA at the probe working concentration of 0.14 mmol/L and 37℃for 20 min,and no positive reaction was found for other common avian pathogens,which was consistent with the results of RT-PCR and chicken embryo isolation and sequencing.The rapid detection method can meet the need of rapid clinical differential diagnosis of H1 subtype AIV,and provide technical support for the rapid detection of grass-roots veterinary field.关键词
H1亚型AIV/等温扩增技术/快速检测/RT-RPA-LFDKey words
H1 subtype AIV/Isothermal amplification technology/Rapid detection/RT-RPA-LFD分类
农业科技引用本文复制引用
曾婷婷,范晴,阮志华,粟永春,李孟,谢芝勋,李丹,谢丽基,罗思思,张民秀,王盛,谢志勤..H1亚型禽流感病毒RT-RPA-LFD快速检测方法的建立及应用[J].江西农业学报,2024,36(3):61-66,6.基金项目
广西重点研发计划项目(桂科AB21076004) （桂科AB21076004）
广西自然科学基金项目(2022GXNSFAA035445、2021GXNSFBA196031) （2022GXNSFAA035445、2021GXNSFBA196031）
广西科技基地和人才专项(桂科AD23049005) （桂科AD23049005）
广西八桂学者(2019-A50). （2019-A50）
