采用离心浓集法提高乙型肝炎病毒表面抗原检出率的可行性研究

Feasibility study on improving detection rate of HBsAg in serum samples by centrifugation-concentration method

目的探讨离心浓集法预处理血清标本对提高乙型肝炎血清学标志物乙型肝炎表面抗原(HBsAg)检出率的可行性研究。方法收集广州市内各大医院2009年52例HBsAg阳性者和疑似乙型肝炎感染者血清标本,分别用酶联免疫吸附试验(ELISA)和化学发光免疫分析法(CLIA)测定HBsAg。再以PEG6000为沉淀剂,将标本进行高速离心浓集(30 000 r/min)25℃,40 min,取沉淀用TE Buffer溶解,采用上述相同方法分别进行测定,比较离心前后其HBsAg的S/CO值与HBsAg定量值的上升情况,用SPSS统计学软件对数据进行非参数检验,分析其差别是否有统计学意义。结果健康对照组与空白对照经高速离心处理前后的HBsAg检测结果差异均无统计学意义(P>0.05),而疑似HBV感染组和HBV感染组患者血清经高速离心预处理后的测定结果均明显高于处理前(P<0.01)。结论离心浓集法预处理对于血清中有低水平乙型肝炎病毒颗粒的感染者有效,血清中的病毒颗粒可通过高速离心处理后得到浓缩,提高临床实验室常规诊断手段的灵敏度的范围,使隐匿型乙型肝炎的检出率有所提高。

Objective To study the possibility of improving detection rate of HBsAg in serum samples by centrifugation-concentration method.Methods 52 serum samples of suspect-HBV patients and HBV infected patients were collected.Then the levels of HBsAg in these samples were tested by ELISA and chemiluminescence immunoassay.After that,the serum samples were treated by polyethylene glycol（PEG）and 40 min＇s ultracentrifugation（30 000 r/min）.The precipitation obtained from each serum was dissolved with TE buffer and the same tests which had been used to test serum samples were carried on them.The anterior-posterior results were compared for statistical significant difference with SPSS 17.0.Results In the HBsAg negative control group,there was no statistical difference in the result of the levels of HBsAg with pretreatment of centrifugation between the two groups（P0.05）.But there was a significant difference in HBV infected group（P0.01）.The level of HBsAg in HBV infected group also increased obviously（P0.01）.Conclusion HBV particle in serum could be concentrated by centrifugation,and which can extend the sensitive range of routine laboratory diagnosed method as well as improve the detection capability of Hepatitis B virus infection.