单扩溶血技术在禽H5N1流感诊断中的应用

Application of single radial hemolysis technigue for diagnosis of influenza A (H5N1)

目的了解单扩溶血技术对禽H5N1流感病毒抗原测定的最佳条件,以便使禽H5N1病毒抗体测定能在普通实验室内进行。方法比较不同浓度,不同品种红细胞,琼脂糖种类和浓度对测定敏感性的影响,以及对该法的敏感性和特异性与微量中和试验进行了比较。结果单扩溶血技术测定最佳条件为:病毒浓度为:1000HA单位/0·1ml浓鸡红细胞;琼脂糖浓度为:1·0%;补体采用后加法。该法的敏感性和特异性不亚于微量中和试验,同时未发现H1N1,尤其N1抗体对H5N1抗体测定有影响。结论单扩溶血技术对禽H5N1病毒抗体测定的敏感性和特异性不亚于微量中和试验,并能在普通实验室对大量血清样本进行测定。

Objective To understand the optimal condition of single radial hemolysis （SRH） for diagnosis of avian influenza A （H5N1） virus in order that SRH could be performed in general laboratories. Methods The effect of different concentration of virus and species of red blood cells, as well as kind and concentration of agarose on testing sensitivity of SRH was determined, Meanwhile the sensitivity and specificity of this method were compared with those of micro-neutralization test. Result The optimal condition of SRH included the viral concentration of 1000 HA units per 0.1 ml packed chicken red blood cells, the agarose concentration of 1.0%, the compliment added into agarose-virus-rbe slides after diffusion of sera. The sensitivity and specificity of SRH were very similar to those of micro-neutralization test. Meanwhile, no cross reaction between antibodies, especially antibodies against N1 antigens, H5N1 and H1N1 viruses was detected. Conclusion The sensitivity and specificity of SRH were very similar to those of micro-neutralization assay. SRH could be performed in normal laboratories and be used for testing large scale serum samples.