Dera-Tomaszewska Bozena, Głośnicka Renata
Akademia Medyczna w Gdańsku, Katedra Mikrobiologii.
Med Dosw Mikrobiol. 2007;59(3):241-50.
The patients' sera had been referred to the National Salmonella Centre for routine Widal serology. Sera were predominately from patients suspected of having been infected with Salmonella Typhi, but also included one serum from patient with typhoid fever who was culture positive for Salmonella Typhi. The immunoblotting procedure using Salmonella Typhi somatic (O=9,12 LPS) and flagellar (H=d) antigens was used for preliminary testing of selected patients sera previously evaluated by Widal agglutination assay as containing different levels of antibodies against O and/or H antigens of Salmonella Typhi. Following Chart et al., immunoblotting reactions were graded between 0 and 3, with 0 indicating an absence of antibody binding, and 3 where antibody binding was readily observed. Sera giving reaction of 2 or 3 were considered to be antibody positive for this study. Positive immunoblotting reaction to O=9,12 LPS antigen was obtained only with the serum of patient with typhoid fever. Presence of specific anti-LPS antibodies was also observed in two other patients' sera diluted 1:50, and in case of one of them also in dilution 1:200, but intensity of antigen-antibody reaction was under positive result criterion. The most other sera positive to O=9,12 antigen in law dilutions (1:50, 1:100) by Widal assay, showed the traces of non-specific reaction by immunoblotting. Presence of positive antigen-antibody reaction was indicated for five sera in dilution 1:50 when tested with the >55 kDa H=d flagellar protein subunit, including the serum of patient with typhoid fever. Only in this serum the high level of specific antibodies was detected also in dilution 1:200, what was not observed in case of the other four, which appeared negative. All the other sera were shown not to contain antibodies to flagella antigen. Although the presented results are preliminary and additional study of more sera of people infected with Salmonella Typhi is needed, it can be concluded after Chart et al., that an immunoblotting procedure incorporating O=9,12 LPS and flagellar H=d antigens is a useful method for providing serological evidence of infection with Salmonella Typhi. In our opinion it can serve as a rapid test for the diagnosis of typhoid fever.
患者血清已被送往国家沙门氏菌中心进行常规肥达氏血清学检测。血清主要来自疑似感染伤寒沙门氏菌的患者,但也包括一份来自伤寒热患者且伤寒沙门氏菌培养呈阳性的血清。使用伤寒沙门氏菌菌体(O = 9,12 LPS)和鞭毛(H = d)抗原的免疫印迹程序,对先前经肥达氏凝集试验评估为含有不同水平针对伤寒沙门氏菌O和/或H抗原抗体的选定患者血清进行初步检测。按照查特等人的方法,免疫印迹反应分为0至3级,0表示无抗体结合,3表示易于观察到抗体结合。在本研究中,反应为2或3的血清被视为抗体阳性。仅伤寒热患者的血清对O = 9,12 LPS抗原获得了阳性免疫印迹反应。在另外两名患者稀释1:50的血清中也观察到了特异性抗LPS抗体的存在,其中一名患者在稀释1:200时也观察到了,但抗原 - 抗体反应强度低于阳性结果标准。其他大多数在肥达氏试验中对O = 9,12抗原低稀释度(1:50, 1:100)呈阳性的血清,通过免疫印迹显示出非特异性反应痕迹。用> 55 kDa H = d鞭毛蛋白亚基检测时,在1:50稀释度下有五份血清显示出阳性抗原 - 抗体反应,包括伤寒热患者的血清。仅在该血清中,在1:200稀释度下也检测到了高水平的特异性抗体,其他四份血清未观察到这种情况,它们呈阴性。所有其他血清均显示不含有针对鞭毛抗原的抗体。尽管所呈现的结果是初步的,并且需要对更多感染伤寒沙门氏菌的人的血清进行进一步研究,但根据查特等人的研究可以得出结论,结合O = 9,12 LPS和鞭毛H = d抗原的免疫印迹程序是提供伤寒沙门氏菌感染血清学证据的有用方法。我们认为它可以作为伤寒热诊断的快速检测方法。