一种用于检测麻风病抗体的酶联免疫吸附测定法的开发。

Development of an ELISA for detection of antibody in leprosy.

作者信息

Douglas J T, Naka S O, Lee J W

出版信息

Int J Lepr Other Mycobact Dis. 1984 Mar;52(1):19-25.

Abstract

An ELISA system was developed for detection of antibodies in leprosy using whole cells of bacteria as an antigen. Whole cells of M. smegmatis, M. vaccae, M. scrofulaceum, M. leprae, C. diphtheriae, and C. xerosis were compared. M. smegmatis was the most reactive against lepromatous sera with OD492 readings 1.5 times and five times higher than the others. In addition, when M. smegmatis were coated to microtiter plates with a volatile ammonium acetate/carbonate buffer and air dried, the antigen coating was found to be three times more reactive than antigen coated with nonvolatile Na borate buffer. Autoclaving M. smegmatis increased the reactivity with lepromatous sera 1.4- to 2.3-fold. M. leprae was found to be 4-10 times more reactive than autoclaved M. smegmatis. Autoclaving M. leprae did not increase reactivity. Antibody titers of some lepromatous sera had endpoint titers of greater than 1:10,000. Both antihuman IgG and antihuman IgA, IgM, and IgG combined conjugates were found to be equally effective in detecting high levels of antibody in patients with multibacillary diseases.

摘要

开发了一种酶联免疫吸附测定（ELISA）系统，以细菌全细胞为抗原检测麻风病抗体。比较了耻垢分枝杆菌、母牛分枝杆菌、瘰疬分枝杆菌、麻风分枝杆菌、白喉棒状杆菌和干燥棒状杆菌的全细胞。耻垢分枝杆菌对瘤型麻风血清反应性最强，其492nm处的光密度（OD）读数分别是其他细菌的1.5倍和5倍。此外，当用挥发性乙酸铵/碳酸盐缓冲液将耻垢分枝杆菌包被到微量滴定板上并风干时，发现抗原包被的反应性是非挥发性硼酸钠缓冲液包被抗原的3倍。对耻垢分枝杆菌进行高压灭菌可使其与瘤型麻风血清的反应性提高1.4至2.3倍。发现麻风分枝杆菌的反应性比经高压灭菌的耻垢分枝杆菌高4至10倍。对麻风分枝杆菌进行高压灭菌不会提高其反应性。一些瘤型麻风血清的抗体效价终点效价大于1:10,000。发现抗人IgG和抗人IgA、IgM和IgG联合偶联物在检测多菌型疾病患者的高水平抗体方面同样有效。