检测鼠疫F1抗原的三种免疫学方法比较

Comparison of three immunological methods in detection of Yersina pestis F1 antigen

目的 比较三种检测鼠疫F1抗原方法用于诊断鼠疫的效果.方法在鼠疫疫源地,收集野生动物的血、肝、脾、淋巴组织标本,酶联免疫吸附试验（ELISA）、反向间接血凝试验（RIHA）、胶体金纸上色谱试验（GICA）三种方法检测鼠疫F1抗原.结果检测鼠疫疫源地内自毙和捕获野生动物脏器浸液标本414份,ELISA和GICA检出的阳性标本相同,阳性率均为5.31%（22/414）,阳性符合率和阴性符合率均为100%.RIHA初试2孔以上凝集186份,复试仅18份阳性,非特异性凝集率高达40.57%（168/414）,阳性率4.35%（18/414）.RIHA与GICA、ELISA的阳性符合率均为81.82%（18/22）,阴性符合率均为98.99%（392/396）;阳性标本的平均滴度,ELISA为1：29.95,RIHA为1：28.68,组间比较差异有统计学意义（t=4.379,P＜0.01）.结论三种方法检测鼠疫F1抗原诊断鼠疫,RIHA结果有定量的意义,非特异性凝集率高,复试工作量大;GICA和ELISA具有相同的特异性和敏感性,GICA结果只有定性意义,ELISA排除了RIHA、GICA两种方法的缺陷,综合了两种方法的优点.

Objective To compare the effect of three methods in diagnosis of plague by detecting of Yersina pestis F1 antigen. Methods In natural foci of plague, wild animal samples, such as blood, liver, spleen,and lymphoid tissue were collected, and the three methods of enzyme linked immunosorbent assay （ELISA）,reverse indirect hemagglutination assay（RIHA） and gold-immunochromatography assay（GICA） were employed to detect F1 antigen of Yersina pestis. Results Total of 414 infused organ samples of natural death and captured wild animals in natural foci of plague were determined. Positive samples detected by GICA and ELISA were the same,the positive rates were 5.31%（22/414）, both positive and negative coincidence rates were consistently 100%. Only 18 samples were positive by retrial in 186 samples with more than 2 holes aggregation by preliminary examination of RIHA, with nonspecific agglutination rate of 40.6% （168/414） and positive rate of 4.35% （18/414）. The positive coincidence rate was 81.82% （18/22） between RIHA with GICA and ELISA, and negative coincidence rate was statistically significant（t = 4.379, P 〈 0.01）. Conclusions ELISA, RIHA and GICA can be used for early diagnosis of plague by detecting F1 antigen. The results of RIHA have quantitative significance, with higher non-specific agglutination rate, and heavy workload of re-examination; GICA and ELISA has the same specificity and sensitivity, but the results of GICA is only qualitative. ELISA excluded the defect of RIHA and GICA, and combines the advantages of both methods.