双抗体夹心斑点酶联法检测轮状病毒的研究

A Double-antibody Sandwich Method of Dot-ELISA for the Detection of Rotavirus

本文报道以本室特制的梳状硝酸纤维素滤膜为载体,采用预先包被特异性抗体的双抗体夹心斑点酶联法检测粪便标本中的轮状病毒。将本法与塑料板 ELISA法(上海市防疫站的试剂盒)对 220例标本检测比较,结果两法符合率为 91.36％,本法阳性率高于板上 ELISA法(P<0.01)。将其中 30份阳性标本进一步作滴度比较,两法相关性好,r=0.8760(P<0.01),本法测得 RV GMT为 119.39,比板上 ELISA法 GMT10.08高 11.8倍,具有显著性差异 P<0.01)。实验还表明本法特异性强、稳定性好、操作简便、结果易于判断。由于预先将抗体包被在梳状膜上(4℃可保存1年),整个检测过程可在二小时之内完成,有助于临床的快速诊断,并易于在基层单位推广应用。

This paper reports a double-antibody sandwich method of Dot-ELISA using comb-like nitro cellulose membrane (NCM) as a solid support for the detection of rotavirus in stool from infantile gastroeneri-tis. The comb-like NCM is coated in advance with the purified antibody.The Comparison between the Dot-ELISA and Conventional ELISA for the detection of RV in 220 stool samples indicated the coincidence rate between two methods was 91. 36% and the positive rate of RV by Dot-ELISA was higher than that by conventional ELISA (P<0. 01) .The sensitivity of two methods for the detection of RV titers in 30 stool samples was compared. The results showed that two methods had a good correlation (r=0. 8760 P<0. 01) and GMT of Dot-ELISA was 11. 8 times higher than that of conventional ELISA, so there was a significant difference (P< 0. 01 ). Meanwhile Dot-ELISA was specific, simple, rapid, reproducible and comb-like NCM Coated with antibody Could be preserved at4 C at least for one year. Therefore whole assay canbe completed with 2 hours. It is suitable for the rapid diagnosis of clinical laboratory.