甘肃医药
甘肅醫藥
감숙의약
Gansu Medical Journal
2014年
6期
401-403
,共3页
AllgloTM探针%白细胞介素-1%单核苷酸多态性
AllgloTM探針%白細胞介素-1%單覈苷痠多態性
AllgloTM탐침%백세포개소-1%단핵감산다태성
AllgloTM probe%IL-1%SNP
目的：探讨AllgloTM探针检测IL-1单核苷酸多态性的实用性。方法：采用AllgloTM 探针实时荧光PCR对IL-1β基因SNP进行基因分型。设计其3954C>T位点AllgloTM探针序列和PCR扩增引物，按照PCR产物特异性和扩增效率，对退火温度、引物浓度等条件进行优化。结果：结果显示AllgloTM探针实时荧光PCR检测体系最适引物浓度为0.4μM，退化温度为58.9℃两步法PCR。在同一条件体系下，检测100例健康体检者IL-1β基因3954C>T位点，并进行基因分型，测序验证其分型结果。结论：AllgloTM探针实时荧光PCR检测IL-1β基因SNP是一种值得推广的高通量、低成本、常规化的基因分型方法。
目的：探討AllgloTM探針檢測IL-1單覈苷痠多態性的實用性。方法：採用AllgloTM 探針實時熒光PCR對IL-1β基因SNP進行基因分型。設計其3954C>T位點AllgloTM探針序列和PCR擴增引物，按照PCR產物特異性和擴增效率，對退火溫度、引物濃度等條件進行優化。結果：結果顯示AllgloTM探針實時熒光PCR檢測體繫最適引物濃度為0.4μM，退化溫度為58.9℃兩步法PCR。在同一條件體繫下，檢測100例健康體檢者IL-1β基因3954C>T位點，併進行基因分型，測序驗證其分型結果。結論：AllgloTM探針實時熒光PCR檢測IL-1β基因SNP是一種值得推廣的高通量、低成本、常規化的基因分型方法。
목적：탐토AllgloTM탐침검측IL-1단핵감산다태성적실용성。방법：채용AllgloTM 탐침실시형광PCR대IL-1β기인SNP진행기인분형。설계기3954C>T위점AllgloTM탐침서렬화PCR확증인물，안조PCR산물특이성화확증효솔，대퇴화온도、인물농도등조건진행우화。결과：결과현시AllgloTM탐침실시형광PCR검측체계최괄인물농도위0.4μM，퇴화온도위58.9℃량보법PCR。재동일조건체계하，검측100례건강체검자IL-1β기인3954C>T위점，병진행기인분형，측서험증기분형결과。결론：AllgloTM탐침실시형광PCR검측IL-1β기인SNP시일충치득추엄적고통량、저성본、상규화적기인분형방법。
Objective:To approach the practicability of the AllgloTM probe for the typing of IL-1 promoter . Methods: The genotyping for IL-1β promoter SNP was performed by the real-time Q-PCR with AllgloTM probe. We designed PCR primers and AllgloTM probe so as to detect the SNP site 3954C>T of IL-1β promoter,and optimized the concentration of primers and anneal temperature in accordance with PCR amplification efficiency and production specificity. Results: We successfully established the AllgloTM probe real-time Q-PCR detecting system, and in this two-step asymmetric PCR method, the primer density and the annealing temperature on the approximation PCR instrument were 0.4μM,58.9℃,respectively.We got the same genotyping by the system and sequenced in 100 healthy people detecting the site 3954C>T of IL-1β gene. Conclusion:The use of AllgloTM probe provides an attractive alternative for genotyping and detection of IL-1β gene SNP,which deserves to spread for its high-flux, low cost and convention.
