用于检测人抗炭疽毒素免疫球蛋白G抗体的多重荧光共价微球免疫测定法与酶联免疫吸附测定法的比较。
Comparison of a multiplexed fluorescent covalent microsphere immunoassay and an enzyme-linked immunosorbent assay for measurement of human immunoglobulin G antibodies to anthrax toxins.
作者信息
Biagini Raymond E, Sammons Deborah L, Smith Jerome P, MacKenzie Barbara A, Striley Cynthia A F, Semenova Vera, Steward-Clark Evelen, Stamey Karen, Freeman Alison E, Quinn Conrad P, Snawder John E
机构信息
Biological Monitoring Laboratory Section, Biomonitoring and Health Assessment Branch, National Institute for Occupational Safety and Health, Cincinnati, Ohio, USA.
出版信息
Clin Diagn Lab Immunol. 2004 Jan;11(1):50-5. doi: 10.1128/cdli.11.1.50-55.2004.
Recently, the Centers for Disease Control and Prevention reported an accurate, sensitive, specific, reproducible, and quantitative enzyme-linked immunosorbent assay (ELISA) for immunoglobulin G (IgG) antibodies to Bacillus anthracis protective antigen (PA) in human serum (C. P. Quinn, V. A. Semenova, C. M. Elie et al., Emerg. Infect. Dis. 8:1103-1110, 2002). The ELISA had a minimum detectable concentration (MDC) of 0.06 microgram/ml, which, when dilution adjusted, yielded a whole-serum MDC of 3.0 micro g of anti-PA IgG per ml. The reliable detection limit (RDL) was 0.09 microgram/ml, while the dynamic range was 0.06 to 1.7 microgram/ml. The diagnostic sensitivity of the assay was 97.6% and the diagnostic specificity was 94.2% for clinically verified cases of anthrax. A competitive inhibition anti-PA IgG ELISA was also developed to enhance the diagnostic specificity to 100%. We report a newly developed fluorescence covalent microbead immunosorbent assay (FCMIA) for B. anthracis PA which was Luminex xMap technology. The FCMIA MDC was 0.006 microgram of anti-PA IgG per ml, the RDL was 0.016 microgram/ml, and the whole-serum equivalent MDC was 1.5 micrograms/ml. The dynamic range was 0.006 to 6.8 microgram/ml. Using this system, we analyzed 20 serum samples for anti-PA IgG and compared our results to those measured by ELISA in a double-masked analysis. The two methods had a high positive correlation (r2 = 0.852; P < 0.001). The FCMIA appears to have benefits over the ELISA for the measurement of anti-PA IgG, including greater sensitivity and speed, enhanced dynamic range and reagent stability, the use of smaller sample volumes, and the ability to be multiplexed (measurement of more than one analyte simultaneously), as evidenced by the multiplexed measurement in the present report of anti-PA and anti-lethal factor IgG in serum from a confirmed clinical anthrax infection.
最近，美国疾病控制与预防中心报告了一种用于检测人血清中抗炭疽芽孢杆菌保护性抗原（PA）免疫球蛋白G（IgG）抗体的准确、灵敏、特异、可重复且定量的酶联免疫吸附测定（ELISA）方法（C.P.奎因、V.A.谢苗诺娃、C.M.伊利等人，《新发传染病》8:1103 - 1110，2002年）。该ELISA的最低检测浓度（MDC）为0.06微克/毫升，经稀释调整后，全血清MDC为每毫升3.0微克抗PA IgG。可靠检测限（RDL）为0.09微克/毫升，动态范围为0.06至1.7微克/毫升。对于经临床确诊的炭疽病例，该检测方法的诊断敏感性为97.6%，诊断特异性为94.2%。还开发了一种竞争性抑制抗PA IgG ELISA，以将诊断特异性提高到100%。我们报告了一种新开发的用于炭疽芽孢杆菌PA的荧光共价微珠免疫吸附测定（FCMIA），它采用了Luminex xMap技术。FCMIA的MDC为每毫升0.006微克抗PA IgG，RDL为0.016微克/毫升，全血清等效MDC为1.5微克/毫升。动态范围为0.006至6.8微克/毫升。使用该系统，我们分析了20份血清样本中的抗PA IgG，并在双盲分析中将我们的结果与ELISA测量结果进行了比较。两种方法具有高度正相关性（r2 = 0.852；P < 0.001）。在检测抗PA IgG方面，FCMIA似乎比ELISA更具优势，包括更高的灵敏度和速度、更大的动态范围和试剂稳定性、使用更小的样本体积以及能够进行多重检测（同时测量多种分析物），本报告中对确诊临床炭疽感染患者血清中的抗PA和抗致死因子IgG进行多重检测就证明了这一点。
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