微反应板酶联夹心杂交定量检测乙型肝炎病毒基因组

目的 建立一种在微量反应板上进行乙型肝炎病毒（ＨＢＶ）基因杂交定量分析的技术。方法 利用人工合成的多组寡核苷酸探针与乙型肝炎病毒全基因组在微量ＤＮＡ结合板上进行杂交反应,并利用自制的标记放大探针检测杂交信号,信号值以Ａ值标示,与标准曲线比较后,可求得待测标本中ＨＢＶ基因含量。结果 （１）本方法可直接从血清标本中对ＨＢＶ定量,可定量范围为２０ｐｇ－２０ｎｇ／ｍｌ;（２）不与人类基因组及其它病毒或细菌

A method for quantification of hepatitis B Virus genome by enzyme linked solid phase sandwich hybridization

Objective To develop a method for quantitative analysis of hepatitis B virus (HBV) genome on microtitre plates. Methods Synthesized oligonucleotides (probes) was utilized to hybridize with HBV genome and hybridized complex was bind to DNA BIND plate. The hybridizing signals were detected and amplified with a signal amplifier which was previously prepared in our lab. With the comparison of A value of sample dish with a standard curve, the amount of HBV genome in samples can be quantified. Results (1) This method can be used to quantitatively analyze HBV genome directly from serum samples with the detectable range from 20pg to 20ng per milliliter. (2) Probes used in our system would not bind to non specific genes. (3) Whole process of quantification can be finished within about 5 hours. Conclusion The technique of enzyme linked solid phase sandwich hybridization is a sensitive, specific and simple method, which can be used for quantitative analysis of HBV genome.