Naidu A K, Gogate A
Department of Microbiology, Lokmanya Tilak Municipal Medical College, Sion, Bombay.
Indian J Pathol Microbiol. 1997 Oct;40(4):531-8.
A one step competitive Enzyme-Linked Immunosorbent assay (ELISA) method was developed to detect mycobacterial antigen in cerebrospinal fluid (CSF) for the diagnosis of tuberculous meningitis and compared with a standard competitive ELISA method. Indigenously prepared soluble extract of Mycobacterium tuberculosis H37 Rv was used as antigen. The study was conducted using CSF of 230 clinically diagnosed cases of tuberculous meningit is and 208 control subjects. A cutoff value of 0.57 ng/ml by the one step ELISA and 0.5 ng/ml by the standard ELISA method were determined. The specificity of both methods were 100% and positivity was 68.26% and 70.43% respectively. A follow up study was conducted in 63 cases at various interval of time after starting anti-tubercular therapy i.e. at 3 weeks (63 cases), 6 weeks (27 cases) and > or = 4-12 months (13 cases). It was observed that antigen levels decreased gradually, but were much above the cutoff range. Indigenously prepared antigen was compared with antigen prepared in other laboratories and standard molecular weight markers using SDS PAGE (Sodium Do-decyl Sulphate Polycrylamide Gel Electrophoresis).
开发了一种一步竞争性酶联免疫吸附测定(ELISA)方法,用于检测脑脊液(CSF)中的分枝杆菌抗原,以诊断结核性脑膜炎,并与标准竞争性ELISA方法进行比较。使用国产结核分枝杆菌H37 Rv可溶性提取物作为抗原。该研究使用了230例临床诊断为结核性脑膜炎的病例的脑脊液和208例对照受试者。确定一步ELISA的临界值为0.57 ng/ml,标准ELISA方法的临界值为0.5 ng/ml。两种方法的特异性均为100%,阳性率分别为68.26%和70.43%。对63例患者在开始抗结核治疗后的不同时间间隔进行了随访研究,即3周(63例)、6周(27例)和≥4至12个月(13例)。观察到抗原水平逐渐下降,但仍远高于临界范围。使用十二烷基硫酸钠聚丙烯酰胺凝胶电泳(SDS PAGE)将国产抗原与其他实验室制备的抗原和标准分子量标记物进行了比较。