It is known that real-time PCR gives exponential signal amplification and real-time detection. Under optimal conditions (100% efficiency) a 10-fold increase in the amount of DNA template is associated with a decrease in Cq value by a factor of 3.4. IPCR-based assays are therefore especially useful for detection of target antigens at large quantitative differences. In contrast, standard ELISA gives linear signal amplification and end point detection and, therefore, suits better for detection of smaller

differences at lower range of concentrations; meaningful calibration curves for ELISA span usually selleck chemicals two orders of magnitude or less. Fifth, the labor requirement of the assays must also be taken into consideration. As shown in Fig. 1, Nano-iPCR based assays are less laborious because they have fever steps than iPCR or ELISA. Once the probes (functionalized Au-NPs) are prepared they can be stored for several months and used immediately for easy quantification of the learn more antigen. Our primary intention was to develop

an assay for detection of cytokines in serum-supplemented cell culture media. Nano-iPCR performed in TopYield strips with master mixes covered with oil film and transparent foil offers a simple and robust assay for rapid detection of IL-3 and SCF using commercially available antibodies and their biotinylated forms. The binding of antibody and thiolated oligonucleotide template to Au-NPs is an easy method of how to combine antibodies and oligonucleotides into a complex suitable for the assays. The assay can be used for quantification of other ligands, provided good monoclonal or polyclonal antibodies and their biotinylated forms are available. In conclusion, Nano-iPCR assay shows enhanced sensitivity and wider dynamic range than ELISA and is easier to perform than iPCR. It can be expected that further improvement of the Nano-iPCR assays, namely introduction of better detection probes with higher ligand specificity and lower nonspecific binding will advance the application of these tests for routine detection of

cytokines as well as other ligands. Synthetically prepared tailored DNA or RNA aptamers (Jayasena, 1999 and Khati, 2010) and tailored recombinant binding proteins (Binz et al., 2005) could provide such probes. The authors do not have a commercial or other association Methocarbamol that might pose a conflict of interest. This work was supported by project KAN200520701 and M200520901 from Academy of Sciences of the Czech Republic, 1M6837805001 (Center of Molecular and Cellular Immunology) and LC-545 from Ministry of Education, Youth and Sports of the Czech Republic, grants 204/05/H023, 301/09/1826 and P302/10/1759 from the Grant Agency of the Czech Republic, and Institutional projectAVOZ50520514. “
“The authors regret the UC MEXUS-CONACYT Postdoctoral Fellowship Program was missing from the acknowledgments in the original publication of this article.