CAJ | 학술논문

高分辨率熔解曲线技术（ High resolution melting，HRM）是近年发展起来的基于实时荧光定量技术分析核苷酸突变而进行基因分型的一种新型技术。比较了几种动物疫病病原（猪瘟病毒、禽白血病病毒、鸡艾美耳球虫株）的不同扩增片段（ C202、ALV450、En573和C965）、不同荧光染料（ LC Green、SYTO9和SYBR GreenⅠ）和不同仪器[LightScanner96（Idaho）， LightCycler480（Roche）和Rotor－Gene Q（Qiagen）]在HRM基因分型上的异同，揭示了HRM技术对病原体进行分型时，不同扩增片段和不同性质荧光染料对其分型结果影响较大：拥有多个熔解区间的核酸序列比单独1个SNP位点特异性高，更适合于分型；核酸相邻熔解区间Tm差值影响实际HRM分型结果；饱和荧光染料（LC Green和SYTO9）比非饱和染料（SYBR GreenⅠ）分辨率更高，更适合用于HRM分析，并且不同浓度荧光染料对Tm值有一定的影响；不同HRM仪器之间HRM结果无明显差异。
고분변솔용해곡선기술（ High resolution melting，HRM）시근년발전기래적기우실시형광정량기술분석핵감산돌변이진행기인분형적일충신형기술。비교료궤충동물역병병원（저온병독、금백혈병병독、계애미이구충주）적불동확증편단（ C202、ALV450、En573화C965）、불동형광염료（ LC Green、SYTO9화SYBR GreenⅠ）화불동의기[LightScanner96（Idaho）， LightCycler480（Roche）화Rotor－Gene Q（Qiagen）]재HRM기인분형상적이동，게시료HRM기술대병원체진행분형시，불동확증편단화불동성질형광염료대기분형결과영향교대：옹유다개용해구간적핵산서렬비단독1개SNP위점특이성고，경괄합우분형；핵산상린용해구간Tm차치영향실제HRM분형결과；포화형광염료（LC Green화SYTO9）비비포화염료（SYBR GreenⅠ）분변솔경고，경괄합용우HRM분석，병차불동농도형광염료대Tm치유일정적영향；불동HRM의기지간HRM결과무명현차이。
High resolution melting curve ( HRM ) technology is a new type of technology for genotyping and mutation scanning developing in recent years based on the technology of real?time fluorescence quantitative PCR.This study compared several causative agents of animal diseases,including Classical swine fever virus/Ei?meria species/Avain leukemia virus.The effects of different amplified regions of pathogens’ genomes(C202/ALV450/ En573 / C965),different fluorescent dyes(SYBR GreenⅠ/ LC Green / SYTO9)and different instruments(LightScanner96(Idaho) / LightCycler480(Roche) / Rotor?Gene Q(Qiagen)) on the HRM?based genotyping.Theexperimental data showed that HRM analysis was mainly affected by different amplified fragments and fluorescentdyes with different properties:nucleic acids with multiple melting range had higher specificity than that of singlenucleotide polymorphism(SNP)and were more suitable for differentiation analysis; difference in Tm on multiplemelting range influenced HRM results; saturated fluorescent dyes(LC Green and SYTO9) had higher resolutionthan unsaturated dye(SYBR GreenⅠ); concentration of fluorescent dye had a greater influence on the value ofTm; there was no significant difference in the HRM results from different PCR?HRM analyzers.