以rCFP10-ESAT6融合蛋白为抗原的ELISA法鉴别Mtb和BCG致敏豚鼠的实验研究

Application of ELISA based on rCFP10 - ESAT6 fusion protein on guinea pigs sensitized by M. tuberculoeie and BCG

目的以rCFP10-ESAT6融合蛋白为抗原，用ELISA法评价其对Mtb和BCG致敏豚鼠的鉴别意义。方法分剐用灭活H37Rv和BEG活菌致敏豚鼠，4周后，以rCFP10-ESAT6及PPD为包被抗原，用ELISA法检测每组豚鼠血清中的结核菌IgG抗体。结果以10只NS对照豚鼠OD490＋2s为正常界限值，rCFP10-ESAT6检测Mtb致敏豚鼠的特异性和敏感性分别为95．24％（20／21）、100％（11／11），PPD检测Mtb致敏脬鼠的特异性和敏感性分别为47．62％（10／21）、100％（11／11），以rCFP10-ESAT6为抗原的阳性值（1．563±0．490）与阴性值（0．166±0．085）的差异比PPD为抗原的阳性值（0．629±0．117）与阴性值（0．206±0．053）的差异更显著。结论以rCFP10-ESAT6为包被抗原的ELISA法能特异区分豚鼠因H37Rv致敏和BCG免疫后产生的抗结核菌抗体。

Objective To evaluate the application of rCFP10 - ESAT6 in ELISA on guinea pigs sensitized by inactivated H37Rv and BCG, Metheds The guinea pigs were sensitized by H37Rv and GCG respectively. 4 weeks later, the ELISA based on rCFP10 - ESAT6 and PPD was set up to detect the Mtb IgG antibody in sera from guinea pigs. Results The positive cutoff value was OD490 plus 2 standard deviation based on negative control guinea pigs. The specificity and sensitivity of ELISA based on rCFP10 - ESAT6 to identify Mtb - sensitized guinea pigs was 95.25% （20/21） and 100% （ 11/11 ） respectively. The disrepancy between the positive value and negative value based on rCFP10 - ESAT6 was more significant than PPD. Conclusion The ELISA based on rCFP10 - ESAT6 can distinguish the antibody induced by H37Rv and BCG.