Instituto de Salud y Ambiente Del Litoral ISAL, CONICET-UNL, Facultad de Bioquímica y Ciencias Biológicas, Universidad Nacional Del Litoral, Ciudad Universitaria, Ruta Nacional N°168, km 472, CPA S3000ZAA, Santa Fe, Argentina.
Laboratorio de Cultivos Celulares (LCC), Facultad de Bioquímica y Ciencias Biológicas, Universidad Nacional Del Litoral, Ciudad Universitaria, Ruta Nacional N°168, km 472, CPA S3000ZAA, Santa Fe, Argentina.
Anal Biochem. 2019 Apr 1;570:56-61. doi: 10.1016/j.ab.2019.02.007. Epub 2019 Feb 12.
One of the most used formats in inmuno-polymerase chain reaction (IPCR) is known as "Universal" IPCR (signal-generating complexes is based on conjugates of biotinylated DNA, biotinylated IgG and avidin). In the present study, we evaluated the utility of using mono- and bi-biotinylated DNA probes, pre-self-assembled DNA-neutravidin complex, blocking step and glutaraldehyde pretreatment of standard PCR tubes to improve the analytical performance of the hTSH-IPCR assay. The use of pre-self-assembled mono-biotinylated DNA-neutravidin complex enhances both the sensitivity and the reproducibility of the hTSH-IPCR assay, even without blocking step: hTSH-IPCR assay showed an improved limit of detection (LOD: 0.01 μIU/ml), calibration sensitivity (SEN: 2.4) and analytic sensitivity (γ: 9 μIU/ml) in comparison with both a self-made ELISA and a commercial one.
免疫聚合酶链反应（IPCR）中最常用的一种方法称为“通用”IPCR（信号生成复合物基于生物素化 DNA、生物素化 IgG 和亲和素的缀合物）。在本研究中，我们评估了使用单和双生物素化 DNA 探针、预自组装 DNA-链亲和素复合物、阻断步骤和戊二醛预处理标准 PCR 管来提高 hTSH-IPCR 测定分析性能的效用。使用预自组装的单生物素化 DNA-链亲和素复合物可提高 hTSH-IPCR 测定的灵敏度和重现性，即使没有阻断步骤也是如此：与自制 ELISA 和商业 ELISA 相比，hTSH-IPCR 测定显示出改进的检测限（LOD：0.01 μIU/ml）、校准灵敏度（SEN：2.4）和分析灵敏度（γ：9 μIU/ml）。
