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TaqMan探针结合熔解曲线分析低拷贝HBVDNA含量

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摘要 目的 通过TaqMan探针结合熔解曲线分析的方法,区分低拷贝HBVDNA检测过程中的非特异性产物,对低拷贝HBVDNA进行准确定量.方法 应用TaqMan探针荧光定量PCR检测900例临床标本,对0~1 000copies/ml的标本直接加入1 μl四维杂交液进行熔解曲线分析,采集标准品和样品的熔解曲线熔点Tm值.结果 900例临床样本中,110例定量结果处于低拷贝0~1 000 copies/ml之间,对110例低拷贝样本进行熔解曲线分析,有75例样本与阳性质粒有同一Tm值,熔解曲线峰位于84.85℃;35例样本与阴性对照有同一Tm值,熔解曲线峰位于76.32℃.其中1例样本浓度在0~500 copeies/ml之间,21例在500~800 copeies/ml之间,53例在800~1 000 copeies/ml之间.结论 TaqMan探针方法结合熔解曲线分析能够排除非特异产物对结果的影响,对低拷贝乙肝病毒进行准确定量.
出处 《现代检验医学杂志》 CAS 2010年第4期74-75,共2页 Journal of Modern Laboratory Medicine
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  • 1Lekanne-Deprez RH, Fijnvandraat AC, Ruijter JM, et al.Sensitivity and accuracy of quantitative real-time polymerase chain reaction using SYBR green Ⅰ depends on cDNA synthesis conditions. Anal Biochem, 2002, 307(1): 63-69.
  • 2Brownie J, Shawcross S, Theaker J, et al. The elimination of primer-dimer accumulation in PCR. Nucleic Acids Res,1997, 25(16): 3235-3241.
  • 3Ball TB, Plummer FA, HayGlass KT. Improved mRNA quantitation in LightCycler RT-PCR. Int Arch Allergy Immunol. 2003 Jan, 130(1): 82-6.
  • 4Karsai A, Muller S, Platz S, et al. Evaluation of a homemade SYBR green Ⅰ reaction mixture for real-time PCR quantification of gene expression. Biotechniques, 2002, 32(4):790-792, 794-796.
  • 5Jung M, Muche JM, Lukowsky A, et al. Dimethyl sulfoxide as additive in ready-to-use reaction mixtures for realtime polymerase chain reaction analysis with SYBR Green Ⅰ dye. Anal Biochem, 2001, 289(2): 292-295.
  • 6Teo IA, Choi JW, Morlese J, et al. LightCycler qPCR optimisation for low copy number target DNA. J Immunol Methods, 2002, 270(1): 119-133.
  • 7Sturzenbaum SR. Transfer RNA reduces the formation of primer artifacts during quantitative PCR. Biotechniques,1999, 27(1): 50-52.
  • 8Vandesompele J, De-Paepe A, Speleman F. Elimination of primer-dimer artifacts and genomic coamplification using a two-step SYBR green Ⅰ real-time RT-PCR. Anal Biochem,2002 , 303(1): 95-98.
  • 9Peters IR, Helps CR, Hall EJ, et al. Real-time RT-PCR:considerations for efficient and sensitive assay design. J Immunol Methods, 2004, 286(1-2): 203-217.

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