地高辛标记寡核苷酸探针对慢性乙肝患者前 C基因突变的检测

DIGOXIN TAGGED OLIGONUCLEOTIDE PROBES DETECT CHRONIC HEPATITIS B PATIENTS WITH PRECORE REGION GENE MUTATION

目的 研究慢性乙肝患者乙肝病毒（ＨＢＶ）前Ｃ区突变情况。方法 用多聚酶链反应（ＰＣＲ）结合地高辛标记的寡核苷酸探针Ｍ０（野毒株）、Ｍ１（１８９８位变异株）、Ｍ２（１８９８／１９０１位变异株）对１６５例慢性乙肝患者前Ｃ区突变情况进行检测。结果 其中１２例ＨＢｓＡｇ（＋），ＨＢｅＡｇ（＋），抗ＨＢｅ（－），抗ＨＢｃ（＋）患者，ＨＢＶＤＮＡ阳性率为１００％，突变株检出率为８．３％。１５３例ＨＢｓＡｇ（＋），ＨＢｅＡｇ（－），抗ＨＢｅ（＋），抗ＨＢｃ（＋）患者，ＨＢＶＤＮＡ总阳性率为１６．３％，突变株总检出率为８４％。结论 前Ｃ基因突变株主要存在于抗ＨＢｅ阳性的慢性乙肝患者体内，ＰＣＲ结合地高辛标记的寡核苷酸探针杂交技术，操作简便快捷，利于临床推广应用。

Objective To study HBV precore region mutation in chronic hepatitis B patients of Shandong province.Method By using digoxin tagged oligonucleotide probes M 0(wild type),M 1(mutated at nt 1898),M 2(mutated at nt 1898/1901) to detect the PCR products of HBV pre C and C gene region in 165 cases of chronic hepatitis B patients in Shandong province.Result In 12 cases of patients with HBsAg(+),HBeAg(+),anti HBe(-),anti HBc(+),the positive rate of HBV DNA was 100%( 12/12),the rate of precore mutations was only 8.3%( 1/12).In 153 cases of patients with HBsAg(+),HBeAg(-),anti HBe(+),anti HBc(+),the positive rate of HBV DNA was 16.3%(25/153),the rate of precore mutations was 84%(21/25).Conclusion These results suggest that precore mutations mainly prevail in chronic hepatitis B patients with serum HBeAg(-),anti HBe(+),the method of using digoxin tagged oligonucleotide probes to detect the precore mutated region is easy and simple,it could be applied to clinical practice widely.