粪便中乙肝病毒的定量检测及其与乙肝病毒标志物的相关性研究

Study on relationship among hepatitis B virus markers and content of HBV-DNA in feces of patients with hepatitis B virus infection

采集50例乙型肝炎未并发消化道出血患者的血液和粪便标本,采用酶联免疫法检测粪便和血清中乙肝病毒标志物(HBVM),QIAmp DNA Stool kit试剂盒法提取粪便总DNA,巢式PCR定性检测粪便中乙肝病毒DNA(HBVDNA),实时荧光定量PCR(FQ-PCR)定量检测血清和粪便中HBV-DNA。根据血清HBVM阳性的不同组合(感染模式)将50例患者分成3组,粪便乙肝表面抗原(HBsAg)阳性37例,血清HBsAg阳性46例,两者检出率差异有统计学意义(P<0.05),1例血清阴性者,粪便中HBsAg阳性。FQ-PCR定量检测血清和粪便HBV-DNA含量分别为4.35±1.49和4.50±1.59。血清HBV-DNA检测,30例阳性(60%),粪便中HBV-DNA共检测出31例阳性(62%)。

We collected the samples offeces and serum for all patients. HBVM in serum and feces were determined with ELISA. Using QIAmp DNA Stool kit to extract human fecal DNA. FQ-PCR were utilized for analysis the content of HBV-DNA in feces and serum while it was detected qualitatively by nested-PCR. Depending on the combination of HBVM, 50 patients were divided into three groups. Hepatitis B surface antigen was positive in 37 cases of stool and 46 cases of serum, both the detection rates were significant （ P 〈0.05 ） . FQ-PCR quantitative detection of HBV-DNA in serum and fecal contents were 4.35 ±1.49 and 4.50 ±1.59 . The positive rates of HBsAg were 74%, 92% in feces and serum, respectively. HBsAg was found in feces in one patient without HBsAg in serum. In 50 patients,the positive rates of HBV-DNA were 62%, 60% in feces and serum,respectively.