应用酶联免疫斑点试验检测入伍新兵结核潜伏感染

Investigation on latent tuberculosis infection in the new recruits with enzyme-linked immunospot assay

目的 应用酶联免疫斑点试验（ELISPOT）检测入伍新兵结核潜伏感染情况，评价ELISPOT在检测结核潜伏感染中的价值。方法以结核菌素纯蛋白衍生物（PPD）皮肤试验为对照，应用ELISPOT试剂盒检测366例2009年驻京部队入伍新兵外周血中分泌结核菌抗原特异性y干扰素（IFN-y）的T淋巴细胞数。对PPD和ELISPOT均为阴性的入伍新兵接种卡介苗，10个月后再做PPD皮肤试验和ELISPOT，比较结果。结果366例入伍新兵中，PPD皮肤试验和ELISPOT阳性率分别为44．81％和31．69％。202例PPD皮肤试验阴性和164例PPD皮肤试验阳性者中，分别有53例（26．24％）和63例（38．41％）ELISPOT阳性，两者的一致率为57．92％（212／366），两者的检测结果差异具有统计学意义（X^2=14．34，P〈0．001）。在接种过卡介苗者中，PPD皮肤试验阳性率为58．53％（127／217），ELISPOT阳性率为29．1）3％（63／217），斑点形成细胞数为32．44±26．52；在未接种卡介苗者中，PPD皮肤试验阳性率为24．83％（37／149），ELISPOT阳性率为35．57％（53／149），斑点形成细胞数为41．81±30．48。110例PPD和ELISPOT均为阴性的人伍新兵接种卡介苗1（1个月后，PPD皮肤试验阳转率为78．18％，而ELISPOT检测均为阴性。结论ELISPOT具有较高的特异性和敏感性，能真实反映入伍新兵的结核潜伏感染情况，可推广应用。

Objective To study the latent tuberculosis infection （LTBI） in the new recruits with enzyme-linked immunospot assay （ELISPOT）, and evaluate the value of ELISPOT on the detection of LTBI. Methods A total of 366 new recruits were intradermally injected with purified protein derivative （PPD） , and detected with ELISPOT assay with recombinant CFP-10/ESAT-6 fusion protein （rCFP 10/ESAT-6） as a stimulus. New recruits who were ELISPOT-and PPD-negative were vaccinated by intradermal injection with Bacillus Calmette-Guerin （BCG） vaecine, they were detected by PPD skin test and ELISPOT assay again 10 months later. Results Among 366 new recruits, the positive rate of PPD skin test and ELISPOT assay was 44. 81%and 31.69%, respectively. Of 202 PPDnegative and 164 PPD-positive new recruits, 53 （26.24 %） and 63 （38.41%） were ELISPOT positive, respectively, overall consistency between two tests was 57. 92% （212/366）, the difference was statistically significant（p（X^2 = 14. 34,P〈0. 001 ）. Among new recruits with BCG vaccination , PPD- and ELISPOT-positive rate was 58. 53% （127/217） and 29. 03% （63/217） respectively, spot forming cell （SFC） were 32. 44 ± 26. 52; Among new recruits without BCG vaccination , PPD- and ELISPOT-positive rate was 24. 83% （37/149） and 35.57M （53/149） respeetively, SFC were 41.81 ± 30. 48. 78. 180/oo of 110 new recruits vaccinated bv BCG vaccine 10 months later were PPD-positive, but all of they were ELISPOT negative. Conclusion ELISPOT technique has high specificity and sensitivity for screening TB infection.