Houghton Raymond L, Lodes Michael J, Dillon Davin C, Reynolds Lisa D, Day Craig H, McNeill Patricia D, Hendrickson Ronald C, Skeiky Yasir A W, Sampaio Diana P, Badaro Roberto, Lyashchenko Konstantin P, Reed Steven G
Corixa Corporation, Seattle, Washington 98104, USA.
Clin Diagn Lab Immunol. 2002 Jul;9(4):883-91. doi: 10.1128/cdli.9.4.883-891.2002.
Screening of genomic expression libraries from Mycobacterium tuberculosis with sera from tuberculosis (TB) patients or rabbit antiserum to M. tuberculosis led to the identification of novel antigens capable of detecting specific antibodies to M. tuberculosis. Three antigens, Mtb11 (also known as CFP-10), Mtb8, and Mtb48, were tested together with the previously reported 38-kDa protein, in an enzyme-linked immunosorbent assay (ELISA) to detect antibodies in TB patients. These four proteins were also produced as a genetically fused polyprotein, which was tested with two additional antigens, DPEP (also known as MPT32) and Mtb81. Sera from individuals with pulmonary and extrapulmonary TB, human immunodeficiency virus (HIV)-TB coinfections, and purified protein derivative (PPD)-positive and PPD-negative status with no evidence of disease were tested. In samples from HIV-negative individuals, the ELISA detected antibodies in >80% of smear-positive individuals and >60% smear-negative individuals, with a specificity of approximately 98%. For this group, smears detected 81.6% but a combination of smear and ELISA had a sensitivity of approximately 93%. The antigen combination detected a significant number of HIV-TB coinfections as well as antibodies in patients with extrapulmonary infections. Improved reactivity in the HIV-TB group was observed by including the antigen Mtb81 that was identified by proteomics. The data indicate that the use of multiple antigens, some of which are in a single polyprotein, can be used to facilitate the development of a highly sensitive test for M. tuberculosis antibody detection.
用结核病（TB）患者的血清或兔抗结核分枝杆菌抗血清筛选结核分枝杆菌的基因组表达文库，从而鉴定出能够检测结核分枝杆菌特异性抗体的新型抗原。在酶联免疫吸附测定（ELISA）中，将三种抗原，即Mtb11（也称为CFP - 10）、Mtb8和Mtb48与先前报道的38 kDa蛋白一起进行检测，以检测结核病患者体内的抗体。这四种蛋白质还被制备成基因融合多蛋白，并与另外两种抗原DPEP（也称为MPT32）和Mtb81一起进行检测。对患有肺结核和肺外结核的个体、人类免疫缺陷病毒（HIV）-结核合并感染患者以及纯化蛋白衍生物（PPD）阳性和PPD阴性且无疾病证据的个体的血清进行了检测。在HIV阴性个体的样本中，ELISA在>80%的涂片阳性个体和>60%的涂片阴性个体中检测到抗体，特异性约为98%。对于该组，涂片检测出81.6%，但涂片和ELISA联合检测的灵敏度约为93%。该抗原组合在大量HIV - 结核合并感染患者以及肺外感染患者中检测到了抗体。通过加入蛋白质组学鉴定出的抗原Mtb81，在HIV - 结核组中观察到了反应性的提高。数据表明，使用多种抗原（其中一些存在于单个多蛋白中）可用于促进开发一种高灵敏度的结核分枝杆菌抗体检测试验。
