ELISA检测唾液中乙型肝炎表面抗原方法的优化与评价

Evaluation of modified ELISA method to detect Hepatitis B surface antigen in saliva specimens

目的探讨优化用于检测唾液中乙型肝炎表面抗原的ELISA法,并对其进行评价。方法对ELISA的不同条件:样品类型、加样体积和孵育温度及时间进行优化,筛选最佳条件;并采用3种不同的临界值计算方法,判断最佳临界值。结果不做任何处理的唾液样品可直接用于检测(P=0.100),50μL唾液样品与100、150μL加样体积检测结果比较差异无统计学意义(P=0.070)。采用25℃孵育18h的优化方法检测的结果和对应血清的检测结果相关性最好,优化后唾液检测乙型肝炎表面抗原方法的特异性和敏感性分别达92.6%和93.6%。结论优化后的ELISA方法在唾液样品的乙型肝炎表面抗原检测中具有潜在的应用价值。

Objective To evaluated a modified ELISA method for detecting the Hepatitis B surface antigen（HBsAg）in saliva samples.Methods Optimized the different conditions,including sample type,sample volume and incubation condition,to screen the best condition.Three different methods to calculate cut-off value was evaluated to screen the most acceptable.Results The nonprocessed saliva sample can be used directly for detection（P=0.100）.Test results of different sample volume was no significant difference（P=0.070）.HBsAg was detected in 44saliva samples out of 47paired positive serum specimens and not detected in 63 saliva samples out of 68matched negative serum samples by the optimized ELISA assay.There was excellent agreement between the results for the serum and saliva specimens and the kappa value was 0.87 for saliva specimens.Using an optimized protocol,the sensitivities and specificities were 93.6%and 92.6%,respectively.Conclusion Our data showed a significant promise for the use of the modified commercial ELISA in saliva sample for Hepatitis B virus infection surveillance.