info:eu-repo/semantics/article
Methodological aspects of Universal immuno-PCR on standard tubes
Fecha
2019-02Registro en:
Abud, Julián Elías; Santamaría, Clarisa Guillermina; Oggero Eberhardt, Marcos Rafael; Rodriguez, Horacio Adolfo; Methodological aspects of Universal immuno-PCR on standard tubes; Academic Press Inc Elsevier Science; Analytical Biochemistry; 570; 2-2019; 56-61
0003-2697
CONICET Digital
CONICET
Autor
Abud, Julián Elías
Santamaría, Clarisa Guillermina
Oggero Eberhardt, Marcos Rafael
Rodriguez, Horacio Adolfo
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.