用血清软琼脂技术研究金黄色葡萄球菌细胞表面蛋白a的存在

Titiek Djannatun
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引用次数: 1

摘要

蛋白A是一些金黄色葡萄球菌(金黄色葡萄球菌)菌株表面的一种特异性蛋白质。葡萄球菌中含有的蛋白A在生物医学中有着广泛的应用。本研究旨在寻找一种简便的方法检测金黄色葡萄球菌蛋白a . 15的存在,以金黄色葡萄球菌cowan I为阳性对照,表皮葡萄球菌和微球菌sp为阴性对照。将细菌接种于10 ml Todd Hewith Broth培养基,Soft Agar (SA)和10 ml血清Soft Agar (SSA)中,用涡旋搅拌,在37℃下孵育18 -24 h。观察生长的菌落形式,并将其分为紧凑型和扩散型菌落。对怀疑含有A蛋白的菌落进行Dot Blot验证。在SSA(添加了SA的兔或鸡血清)中,7株菌落由弥漫性变为致密性,8株菌落仍为弥漫性。斑点斑点试验阳性。蛋白A能与IgG的Fc部分结合,但不能与鸟类的IgY结合。金黄色葡萄球菌与IgG的结合可以通过菌落生长由加入含有IgG的血清前的弥漫性变为血清存在后的致密性来证明。细菌细胞表面的蛋白A与Fc-IgG结合,引起细菌的位阻,在SA中以紧密集落的形式表达。利用哺乳动物血清的SSA可用于区分含A蛋白或不含A蛋白的菌株。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
The Presence of Protein a on the Surface of Staphylococcus Aureus Bacterial Cells Using Serum Soft Agar Techniques
Protein A is a specific proteins on the surface of some  Staphylococcus aureus (S. aureus) strains. Staphylococcus bacteria which ha ve protein A has widely used in the biomedical . Aims of this study  to find a simple method of detecting the presence of S. aureus protein A. 15 S. aureus isolates from field cases, S. aureus cowan I as positive control, Staphylococcus epidermidis dan Micrococcus sp as negatif control were used .  Bacteria was inoculated into 10 ml Todd Hewith Broth medium, Soft Agar (SA) and into 10 ml of serum soft-agar (SSA) , agitated using a vortex and incubated at 37 o C for 18 -24 h . Form of colonies that grew was observed and categorized as compact and diffuse colonies . The c olonies that suspected contain protein A conducted confirmation tests by Dot Blot . 7  isolates showed the change of colony formation from diffuse to compact in SSA (SA added rabbit or chicken serum), and 8 isolates remain diffuse. Dot Blot test positive. Protein A has the ability to bind the Fc fraction of IgG but not the IgY from birds. The binding of S. aureus and IgG could be demonstrated by the change of colony growth from diffuse before the addition of serum containing IgG in SA to compact after the presence of serum. Protein A on the bacterial cell surface bind the Fc-IgG caused steric hinderance of bacteria and expressed as compact colony formation in SA. SSA using mammalian sera can be used to discriminate the bacterial strains with or without protein A.
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