CAJ | 학술논문

为了建立检测 H3N8亚型马流感病毒的RT‐LAMP方法，根据 H3N8亚型马流感病毒 HA基因序列，设计了2对LAM P引物，经过优化反应条件，建立了RT‐LAM P检测 H3N8亚型马流感病毒方法。结果表明，RT‐LAMP方法能够在63℃恒温条件下、75 min内实现目的基因片段的大量特异性扩增，通过荧光显色就可直接用肉眼判断结果。对 H7N7亚型马流感病毒、马动脉炎病毒、马鼻肺炎病毒的核酸无交叉反应，具有较好的特异性；方法的灵敏度比常规RT‐PCR的高10倍；该方法操作简便快速、省时省力，而且灵敏度高、特异性强，对 H3N8亚型马流感病毒的检测具有实际的应用价值。
위료건립검측 H3N8아형마류감병독적RT‐LAMP방법，근거 H3N8아형마류감병독 HA기인서렬，설계료2대LAM P인물，경과우화반응조건，건립료RT‐LAM P검측 H3N8아형마류감병독방법。결과표명，RT‐LAMP방법능구재63℃항온조건하、75 min내실현목적기인편단적대량특이성확증，통과형광현색취가직접용육안판단결과。대 H7N7아형마류감병독、마동맥염병독、마비폐염병독적핵산무교차반응，구유교호적특이성；방법적령민도비상규RT‐PCR적고10배；해방법조작간편쾌속、성시성력，이차령민도고、특이성강，대 H3N8아형마류감병독적검측구유실제적응용개치。
To develop a rapid ,sensitive ,and specific approach for the detection of H 3N8 subtype equine in‐fluenza virus (EIV) ,an RT‐LAMP assay with four pairs of primers designed according to the conserved sequences of EIV HA gene was established in the present study .With optimized reaction condition ,the vi‐ral RNA was amplified within 75 min at 63 ℃ ,then the positive results of RT‐LAMP assay were judged through visible green .The established RT‐LAMP was highly specific and no cross reaction with other e‐quine viruses such as H7N7 subtype EIV ,EAV ,EHV .The sensitivity was 10 times higher than that of conventional RT‐PCR .The method established provided a promising method for the detection of H 3N8 subtype EIV .