Nandakumar K S, Palanivel V, Muthukkaruppan V
Department of Immunology, School of Biological Sciences, Madurai Kamaraj University, India.
Clin Exp Immunol. 1993 Nov;94(2):317-21. doi: 10.1111/j.1365-2249.1993.tb03450.x.
Porins are highly immunogenic outer membrane proteins of Salmonella. Sera from typhoid patients contained a high level of IgG antibodies directed to porins of Salm. typhi. Since porins are highly conserved proteins, anti-porins antibodies both from typhoid patients and healthy normals reacted with porins from several Gram-negative bacteria. Therefore, in order to improve the specificity of detecting Salm. typhi porins-specific antibodies, an inhibition ELISA was developed using enzyme-conjugated MoAbs (MP1 and MPN4) specific to Salm. typhi porins. Sera from typhoid patients with positive haemoculture (16 out of 17) inhibited the binding of MP1 to porins, thus showing a positive test for typhoid, whereas sera from patients with other Gram-negative bacterial infections (n = 7) and from healthy volunteers (66 out of 67) were found to be negative. The sensitivity, specificity, accuracy, positive predictive value and negative predictive value of this assay were 94.1, 98.7, 97.8, 94.1 and 98.7% respectively. The validity of our inhibition ELISA for typhoid was higher than that of the Widal test. The diagnosis of typhoid fever as early as 3 days after the onset of fever, using a single specimen is possible.
孔蛋白是沙门氏菌具有高度免疫原性的外膜蛋白。伤寒患者的血清中含有高水平的针对伤寒沙门氏菌孔蛋白的IgG抗体。由于孔蛋白是高度保守的蛋白质,伤寒患者和健康正常人的抗孔蛋白抗体均能与几种革兰氏阴性菌的孔蛋白发生反应。因此,为了提高检测伤寒沙门氏菌孔蛋白特异性抗体的特异性,开发了一种抑制ELISA,使用针对伤寒沙门氏菌孔蛋白的酶联单克隆抗体(MP1和MPN4)。血培养阳性的伤寒患者血清(17例中有16例)抑制了MP1与孔蛋白的结合,因此伤寒检测呈阳性,而其他革兰氏阴性菌感染患者(n = 7)和健康志愿者(67例中有66例)的血清检测为阴性。该检测方法的灵敏度、特异性、准确性、阳性预测值和阴性预测值分别为94.1%、98.7%、97.8%、94.1%和98.7%。我们的伤寒抑制ELISA的有效性高于肥达试验。使用单个标本有可能在发热开始后3天就诊断出伤寒热。