烟曲霉单克隆抗体的制备与鉴定以及烟曲霉GM抗原捕获方法的建立
发布时间：2018-07-28 09:10
【摘要】：近年来,由于各种医源性因素如广谱抗生素、皮质类固醇激素、免疫抑制剂以及抗肿瘤药物等的广泛使用,器官移植及各种导管介入手术的开展,导致机会性深部真菌感染的发病率明显增高。曲霉菌跃居第二已成为仅次于念珠菌的重要致病性真菌,由曲霉感染引起的侵袭性曲霉病(IA),严重威胁患者的生命,死亡率高达80%甚至到100%,其中烟曲霉感染引起的IA占总IA的90%以上。临床研究表明,IA的早期诊断与预后关系密切。但由于曲霉感染发病隐匿,临床表现缺乏特征性,早期诊断十分困难。血培养、组织活检为曲霉感染实验室诊断的金标准,但培养时间长、阳性率低、操作污染导致的假阳性及组织活检的局限性和创伤性,已不能满足临床的需要。目前,血清学和分子生物学研究已成为人们的关注点,PCR技术在早期感染即可检测到微量的真菌DNA,且可以鉴定到种,对指导临床用药有一定帮助。但在引物设计、临床标本的防污染处理,结果的判断等对实验室和操作人员的要求很高,国内外仍停留在小样本的研究。而血清抗体的检测受限于患者本身在严重免疫抑制状态下,无法产生足够的抗体。而且由于烟曲霉孢子漂浮于空气中,几乎每个健康成人血清中均存在烟曲霉抗体。因此,抗体不能用于早期、特异性的检测。真菌抗原的检测也是目前研究的热点。研究显示,从烟曲霉的提取物中目前至少发现100多种抗原性分子,约有12种被纯化成单一抗原形式。已经证实烟曲霉细胞壁上丰富的半乳糖甘露聚糖(galactomannan GM)是烟曲霉的主要抗原成分,
[Abstract]:In recent years, due to the extensive use of various iatrogenic factors such as broad-spectrum antibiotics, corticosteroids, immunosuppressants and anti-tumor drugs, organ transplantation and various catheter intervention operations have been carried out. The incidence of opportunistic deep fungal infection was significantly increased. Aspergillus has become the second most important pathogenic fungus after candida. The invasive aspergillosis (IA),) caused by aspergillus infection is a serious threat to the life of patients. Mortality was as high as 80% to 100%, of which IA caused by Aspergillus fumigatus accounted for more than 90% of the total IA. Clinical studies have shown that early diagnosis of IA is closely related to prognosis. However, the early diagnosis of aspergillus infection is very difficult because of its hidden incidence and lack of characteristic clinical manifestations. Blood culture and tissue biopsy are the gold standard for laboratory diagnosis of aspergillus infection, but the culture time is long, the positive rate is low, the false positive caused by operation contamination and the limitation and trauma of tissue biopsy can not meet the clinical needs. At present, serology and molecular biology research have become the focus of attention. PCR technology can detect trace fungal DNA in early infection, and can identify species, which is helpful to guide clinical drug use. However, primer design, anti-contamination treatment of clinical specimens and judgment of results are very demanding for laboratory and operator, and they are still in the research of small samples at home and abroad. The detection of serum antibodies is limited by the fact that the patients themselves are unable to produce enough antibodies under severe immunosuppressive conditions. And because Aspergillus fumigatus spores float in the air, almost every healthy adult serum contains Aspergillus fumigatus antibody. Therefore, antibodies cannot be used for early, specific detection. The detection of fungal antigens is also a hot topic at present. Studies show that at least 100 antigenic molecules have been found from the extracts of Aspergillus fumigatus and about 12 of them have been purified into a single antigen form. It has been proved that the abundant galactosmannan (galactomannan GM) on the cell wall of Aspergillus fumigatus is the main antigen of Aspergillus fumigatus.
【学位授予单位】：第一军医大学
【学位级别】：硕士
【学位授予年份】：2005
【分类号】：R392
[Abstract]:In recent years, due to the extensive use of various iatrogenic factors such as broad-spectrum antibiotics, corticosteroids, immunosuppressants and anti-tumor drugs, organ transplantation and various catheter intervention operations have been carried out. The incidence of opportunistic deep fungal infection was significantly increased. Aspergillus has become the second most important pathogenic fungus after candida. The invasive aspergillosis (IA),) caused by aspergillus infection is a serious threat to the life of patients. Mortality was as high as 80% to 100%, of which IA caused by Aspergillus fumigatus accounted for more than 90% of the total IA. Clinical studies have shown that early diagnosis of IA is closely related to prognosis. However, the early diagnosis of aspergillus infection is very difficult because of its hidden incidence and lack of characteristic clinical manifestations. Blood culture and tissue biopsy are the gold standard for laboratory diagnosis of aspergillus infection, but the culture time is long, the positive rate is low, the false positive caused by operation contamination and the limitation and trauma of tissue biopsy can not meet the clinical needs. At present, serology and molecular biology research have become the focus of attention. PCR technology can detect trace fungal DNA in early infection, and can identify species, which is helpful to guide clinical drug use. However, primer design, anti-contamination treatment of clinical specimens and judgment of results are very demanding for laboratory and operator, and they are still in the research of small samples at home and abroad. The detection of serum antibodies is limited by the fact that the patients themselves are unable to produce enough antibodies under severe immunosuppressive conditions. And because Aspergillus fumigatus spores float in the air, almost every healthy adult serum contains Aspergillus fumigatus antibody. Therefore, antibodies cannot be used for early, specific detection. The detection of fungal antigens is also a hot topic at present. Studies show that at least 100 antigenic molecules have been found from the extracts of Aspergillus fumigatus and about 12 of them have been purified into a single antigen form. It has been proved that the abundant galactosmannan (galactomannan GM) on the cell wall of Aspergillus fumigatus is the main antigen of Aspergillus fumigatus.
【学位授予单位】：第一军医大学
【学位级别】：硕士
【学位授予年份】：2005
【分类号】：R392
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