PDF(Pubmed)
Abstract：
UNASSIGNED: The method currently available to diagnose shigellosis is insensitive and has many limitations. Thus, this study was designed to identify specific antigenic protein(s) among the cell surface associated proteins (SAPs) of Shigella that would be valuable in the development of an alternative diagnostic assay for shigellosis, particularly one that could be run using a stool sample rather than serum.
UNASSIGNED: The SAPs of clinical isolates of S. dysenteriae, S. boydii, Shigella flexneri, and S. sonnei were extracted from an overnight culture grown at 37 °C using acidified-glycine extraction methods. Protein profiles were observed by SDS-PAGE. To determine if antibodies specific to certain Shigella SAPs were present in both sera and stool suspensions, Western blot analysis was used to detect the presence of IgA, IgG, and IgM.
UNASSIGNED: Immunoblot analysis revealed that sera from patients infected with S. flexneri recognized 31 proteins. These SAP antigens are recognized by the host humoral response during Shigella infection. Specific antibodies against these antigens were also observed in intestinal secretions of shigellosis patients. Of these 31 S. flexneri proteins, the 35 kDa protein specifically reacted against IgA present in patients\' stool suspensions. Further study illustrated the immunoreactivity of this protein in S. dysenteriae, S. boydii, and S. sonnei. This is the first report that demonstrates the presence of immunoreactive Shigella SAPs in stool suspensions. The SAPSs could be very useful in developing a simple and rapid serodiagnostic assay for shigellosis directly from stool specimens.
UNASSIGNED: The SAPs of clinical isolates of S. dysenteriae, S. boydii, Shigella flexneri, and S. sonnei were extracted from an overnight culture grown at 37 °C using acidified-glycine extraction methods. Protein profiles were observed by SDS-PAGE. To determine if antibodies specific to certain Shigella SAPs were present in both sera and stool suspensions, Western blot analysis was used to detect the presence of IgA, IgG, and IgM.
UNASSIGNED: Immunoblot analysis revealed that sera from patients infected with S. flexneri recognized 31 proteins. These SAP antigens are recognized by the host humoral response during Shigella infection. Specific antibodies against these antigens were also observed in intestinal secretions of shigellosis patients. Of these 31 S. flexneri proteins, the 35 kDa protein specifically reacted against IgA present in patients\' stool suspensions. Further study illustrated the immunoreactivity of this protein in S. dysenteriae, S. boydii, and S. sonnei. This is the first report that demonstrates the presence of immunoreactive Shigella SAPs in stool suspensions. The SAPSs could be very useful in developing a simple and rapid serodiagnostic assay for shigellosis directly from stool specimens.
摘要：
■目前可用于诊断志贺氏菌病的方法不敏感，并且有许多局限性。因此，本研究旨在鉴定志贺氏菌的细胞表面相关蛋白（SAP）中的特定抗原蛋白，这些蛋白在志贺氏菌病的替代诊断试验的开发中具有重要价值。特别是可以使用粪便样本而不是血清的样本。
■痢疾沙门氏菌临床分离株的SAP，S.boydii,福氏志贺氏菌,使用酸化的甘氨酸提取方法从在37°C下生长的过夜培养物中提取S.sonnei。通过SDS-PAGE观察蛋白质谱。为了确定某些志贺氏菌SAP特异性抗体是否存在于血清和粪便悬浮液中，Westernblot分析用于检测IgA的存在，IgG,和IgM。
■免疫印迹分析显示，来自感染福氏链球菌的患者的血清识别31种蛋白质。这些SAP抗原在志贺氏菌感染期间被宿主体液应答识别。在志贺氏菌病患者的肠道分泌物中也观察到针对这些抗原的特异性抗体。在这31种福氏杆菌蛋白中，35kDa蛋白对患者粪便悬浮液中存在的IgA特异性反应。进一步的研究说明了该蛋白在痢疾沙门氏菌中的免疫反应性，S.boydii,还有S.sonnei.这是证明粪便悬浮液中存在免疫反应性志贺氏菌SAP的第一份报告。SAPS对于直接从粪便标本中开发一种简单，快速的志贺氏菌病血清诊断方法非常有用。
■痢疾沙门氏菌临床分离株的SAP，S.boydii,福氏志贺氏菌,使用酸化的甘氨酸提取方法从在37°C下生长的过夜培养物中提取S.sonnei。通过SDS-PAGE观察蛋白质谱。为了确定某些志贺氏菌SAP特异性抗体是否存在于血清和粪便悬浮液中，Westernblot分析用于检测IgA的存在，IgG,和IgM。
■免疫印迹分析显示，来自感染福氏链球菌的患者的血清识别31种蛋白质。这些SAP抗原在志贺氏菌感染期间被宿主体液应答识别。在志贺氏菌病患者的肠道分泌物中也观察到针对这些抗原的特异性抗体。在这31种福氏杆菌蛋白中，35kDa蛋白对患者粪便悬浮液中存在的IgA特异性反应。进一步的研究说明了该蛋白在痢疾沙门氏菌中的免疫反应性，S.boydii,还有S.sonnei.这是证明粪便悬浮液中存在免疫反应性志贺氏菌SAP的第一份报告。SAPS对于直接从粪便标本中开发一种简单，快速的志贺氏菌病血清诊断方法非常有用。
