基于微滴式數(shù)字PCR技術(shù)的甲型流感病毒絕對定量方法的建立及應(yīng)用
發(fā)布時(shí)間：2019-06-21 04:41
【摘要】：數(shù)字PCR(Digital PCR,dPCR)是核酸絕對定量的新方法,然而,基于dPCR的甲型流感病毒(Influenza A virus,FluA)絕對定量方法還未系統(tǒng)建立。本研究首先對微滴式dPCR的退火溫度進(jìn)行了梯度優(yōu)化,確定了dPCR反應(yīng)的最佳退火溫度為64.4℃;利用FluA核酸標(biāo)準(zhǔn)品,確定了微滴式dPCR對FluA的檢測范圍為37.7~8.22"104拷貝/#L,檢測的檢出限為3.77拷貝/反應(yīng)。微滴式dPCR的檢測結(jié)果與標(biāo)準(zhǔn)品拷貝數(shù)的相關(guān)系數(shù)為R2=0.9988,提示該方法檢測結(jié)果具有較高的可信度。用建立好的微滴式dPCR方法可對待測臨床樣本中的FluA進(jìn)行了拷貝數(shù)定量。因此本研究建立了基于微滴式dPCR的FluA絕對定量方法,可有效地對臨床樣本中甲型流感病毒載量進(jìn)行絕對定量,為臨床研究中病毒載量的測定提供了一種技術(shù)。
[Abstract]:Digital PCR (Digital PCR,dPCR) is a new method for absolute quantification of nucleic acid. However, the absolute quantitative method of influenza A virus (Influenza A virus,FluA) based on dPCR has not yet been systematically established. In this study, the annealing temperature of microdrop dPCR was optimized, and the optimum annealing temperature of dPCR reaction was 64.4 鈩，
本文編號：2503796
[Abstract]:Digital PCR (Digital PCR,dPCR) is a new method for absolute quantification of nucleic acid. However, the absolute quantitative method of influenza A virus (Influenza A virus,FluA) based on dPCR has not yet been systematically established. In this study, the annealing temperature of microdrop dPCR was optimized, and the optimum annealing temperature of dPCR reaction was 64.4 鈩，
本文編號：2503796
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