Khalilpour Akbar, Santhanam Amutha, Wei Lee Chun, Saadatnia Geita, Velusamy Nagarajan, Osman Sabariah, Mohamad Ahmad Munir, Noordin Rahmah
Institute for Research in Molecular Medicine, Universiti Sains Malaysia, Minden, Malaysia.
Asian Pac J Cancer Prev. 2013;14(3):1635-42. doi: 10.7314/apjcp.2013.14.3.1635.
Helicobacter pylori antigen was prepared from an isolate from a patient with a duodenal ulcer. Serum samples were obtained from culture-positive H. pylori infected patients with duodenal ulcers, gastric ulcers and gastritis (n=30). As controls, three kinds of sera without detectable H. pylori IgG antibodies were used: 30 from healthy individuals without history of gastric disorders, 30 from patients who were seen in the endoscopy clinic but were H. pylori culture negative and 30 from people with other diseases. OFF-GEL electrophoresis, SDS-PAGE and Western blots of individual serum samples were used to identify protein bands with good sensitivity and specificity when probed with the above sera and HRP-conjugated anti-human IgG. Four H. pylori protein bands showed good (≥ 70%) sensitivity and high specificity (98-100%) towards anti-Helicobacter IgG antibody in culture- positive patients sera and control sera, respectively. The identities of the antigenic proteins were elucidated by mass spectrometry. The relative molecular weights and the identities of the proteins, based on MALDI TOF/ TOF, were as follows: CagI (25 kDa), urease G accessory protein (25 kDa), UreB (63 kDa) and proline/pyrroline- 5-carboxylate dehydrogenase (118 KDa). These identified proteins, singly and/or in combinations, may be useful for diagnosis of H. pylori infection in patients.
幽门螺杆菌抗原是从一名十二指肠溃疡患者的分离株中制备的。血清样本取自经培养证实感染幽门螺杆菌的十二指肠溃疡、胃溃疡和胃炎患者(n = 30)。作为对照,使用了三种未检测到幽门螺杆菌IgG抗体的血清:30份来自无胃部疾病史的健康个体,30份来自在内镜检查门诊就诊但幽门螺杆菌培养阴性的患者,以及30份来自患有其他疾病的人群。对个体血清样本进行离胶电泳、SDS - 聚丙烯酰胺凝胶电泳和蛋白质印迹分析,当用上述血清和辣根过氧化物酶偶联的抗人IgG进行检测时,可识别出具有良好敏感性和特异性的蛋白条带。四条幽门螺杆菌蛋白条带分别对培养阳性患者血清和对照血清中的抗幽门螺杆菌IgG抗体表现出良好(≥70%)的敏感性和高特异性(98 - 100%)。通过质谱分析阐明了抗原蛋白的身份。基于基质辅助激光解吸电离飞行时间/飞行时间质谱(MALDI TOF/TOF),这些蛋白的相对分子量和身份如下:CagI(25 kDa)、脲酶G辅助蛋白(25 kDa)、UreB(63 kDa)和脯氨酸/吡咯啉 - 5 - 羧酸脱氢酶(118 KDa)。这些已鉴定的蛋白单独和/或组合使用,可能有助于诊断患者的幽门螺杆菌感染。