Methodological aspects of Universal immuno-PCR on standard tubes

被引：3

作者：

Abud, J. E. ^{[1
]}

Santamaria, C. G. ^{[1
]}

Oggero, M. ^{[2
]}

Rodriguez, H. A. ^{[1
]}

机构：

[1] Univ Nacl Litoral, CONICET, Fac Bioquim & Ciencias Biol, ISAL, Ruta Nacl 168,Km 472,CPA S3000ZAA, Santa Fe, Argentina

[2] Univ Nacl Litoral, LCC, Fac Bioquim & Ciencias Biol, Ruta Nacl 168,Km 472,CPA S3000ZAA, Santa Fe, Argentina

来源：

ANALYTICAL BIOCHEMISTRY | 2019年 / 570卷

关键词：

Immune-PCR; Human TSH; Diagnostic; Monoclonal antibodies; Immunoassays; CHAIN-REACTION ASSAY; ULTRASENSITIVE DETECTION; PROTEIN; DNA; QUANTIFICATION; NANOSTRUCTURES; SENSITIVITY; FIGURES; DETECT; MERIT;

D O I：

10.1016/j.ab.2019.02.007

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

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 mu IU/ml), calibration sensitivity (SEN: 2.4) and analytic sensitivity (gamma: 9 mu IU/ml(-1)) in comparison with both a self-made ELISA and a commercial one.

引用

页码：56 / 61

页数：6

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