Antigenemia immunoassay for point of care dianostic melioidosis

抗原免疫分析用于诊断类鼻疽病的护理点

• 批准号: 8260262
• 负责人: David P AuCoin
• 金额: $ 28.71万
• 依托单位: UNIVERSITY OF CALIFORNIA-IRVINE
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-05-01 至 2014-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8260262
• 关键词: 3-Dimensional; Acute; Admission activity; Antibiotics; Antibodies; Antigen Targeting; Antigens; Bacteremia; Bacteria; Bacterial Infections; Biological Assay; Blood; Body Fluids; Burkholderia pseudomallei; Categories; Detection; Diagnosis; Disaccharides; Early Diagnosis; Emerging Communicable Diseases; Enzyme-Linked Immunosorbent Assay; Goals; Gold; Human; Human Resources; Immunoassay; Immunological Diagnosis; Infection; Infectious Diseases Research; Laboratory Diagnosis; Lipopolysaccharides; Medical; Melioidosis; Monoclonal Antibodies; Mus; Outcome; Patients; Polymers; Polysaccharides; Process; Proteins; Reporting; Sepsis; Septicemia; Serum; Site; Source; Staging; Structure; biodefense; biothreat; experience; extracellular; falls; human disease; in vivo; microbial; point of care; public health emergency; rapid diagnosis

Status of laboratory diagnosis of melioidosis Currently, culture from multiple body sites is the gold standard for laboratory diagnosis. Culture requires experienced personnel and takes 3-4 days. Levels of bacteremia are very low (~ i CFU/ml). A point-of-care immunoassay for diagnosis of melioidosis could greatly impact patient outcome because a high percentage of patients with acute septicemia die within 24-48 h of admission, and the antibiotics used for empiric treatment of septicemia are not effective for B. pseudomallei. Assays for antibody have limited value due to an absence of antibody in the early stages of acute sepsis and a high level of background antibodies in endemic regions (6). Several studies reported PCR for detection of B. pseudomallei, but the reported lower limit of detection tends to fall above the range for the viable bacteria count in blood during human disease (118). Potential antigen targets for immunodiagnosis of melioidosis Detection of extracellular polysaccharides has been successful for a variety of bacterial infections. There are three candidate antigens for B. pseudomallei: the exopolysaccharide (EPS), the capsular polysaccharide (CPS), and the lipopolysaccharide (LPS). EPS is an unbranched polymer of a repeating tetrasaccharide: [-3)-p-D-Galp2Ac-(i-4)-a-D-Galp-(i-3)-p-D-Galp-(i-5)-|3-Kdo-(2-] (115). CPS is an unbranched homopolymer of [-3)-2-O-acetyl-6-deoxy-p-D-mannoheptopyranose-( i-] (120). LPS is an unbranched polymer of disaccharide repeating units having the basic structure: [-3-)-|3-D-glucopyranose-(i-3)-6-deoxy-a-L-talopyranose-(i-] (120). Melioidosis patients make antibodies to EPS, CPS and LPS, indicating that these antigens are produced in vivo. To date, no B. pseudomallei proteins have been identified as candidates for antigen-targeted diagnosis of melioidosis. The proposed InMAD process has the potential to identify B. pseudomallei antigens shed in vivo and may serve as a rapid broad discovery platform for many microbial threats.

类鼻疽实验室诊断现状 目前，多部位培养是金科玉律 实验室诊断标准。文化需要有经验的人员，需要3-4天。级别 菌血症非常低（~ i CFU/ml）。用于诊断类鼻疽的即时免疫分析可以极大地改善 影响患者的治疗结果，因为大部分急性败血症患者在 24-48 小时内死亡 入院时，用于败血症经验性治疗的抗生素对类鼻疽杆菌无效。 由于急性脓毒症早期阶段缺乏抗体，因此抗体检测的价值有限。 流行地区背景抗体水平较高 (6)。多项研究报道了 PCR 检测 B. 假鼻疽，但报告的检测下限往往低于活细菌的范围 人类疾病期间血液中的计数 (118)。 类鼻疽免疫诊断的潜在抗原靶点 细胞外检测 多糖已成功治疗多种细菌感染。类鼻疽杆菌有三种候选抗原：胞外多糖 (EPS)、荚膜多糖 (CPS) 和脂多糖 (LPS)。 EPS 是重复四糖的无支链聚合物：[-3)-p-D-Galp2Ac-(i-4)-a-D-Galp-(i-3)-p-D-Galp-(i-5)-|3-Kdo-(2-] (115)。CPS 是无支链均聚物 [-3)-2-O-乙酰基-6-脱氧-对-D-吡喃甘露庚糖-( 我-]（120）。 LPS 是二糖重复单元的无支化聚合物，具有以下基本结构： 结构：[-3-)-|3-D-吡喃葡萄糖-(i-3)-6-脱氧-a-L-吡喃糖-(i-] (120)。类鼻疽患者产生针对 EPS、CPS 和 LPS 的抗体，表明这些抗原是在体内产生的。迄今为止，尚未确定类鼻疽伯克霍尔德蛋白可作为抗原靶向诊断的候选者。 类鼻疽。拟议的 InMAD 过程有可能识别体内脱落的类鼻疽伯克氏菌抗原，并可作为许多微生物威胁的快速广泛发现平台。