INVESTIGADORES
OGGERO EBERHARDT Marcos Rafael
artículos
Título:
Methodological aspects of Universal immuno-PCR on standard tubes
Autor/es:
ABUD, J.E.; SANTAMARÍA, C.G.; OGGERO, M.; RODRIGUEZ, H.A.
Revista:
ANALYTICAL BIOCHEMISTRY
Editorial:
ACADEMIC PRESS INC ELSEVIER SCIENCE
Referencias:
Año: 2019 vol. 570 p. 56 - 61
ISSN:
0003-2697
Resumen:
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 −1 ) in comparison with both a self-made ELISA and a commercial one.
