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The above HhCG assays were performed manually using large sample volumes (200 [micro]L) and long incubation times (turnaround time, 1-2 days).
During a 30-min incubation at 37[degrees]C, HhCG in the sample bound to the B152 capture antibody, which in turn bound to the magnetic particles.
The limit of quantification, based on assays of serial dilutions of a sample (HhCG = 2 [micro]g/L) with the CV calculated from 10 observations at each dilution, was 0.2 [micro]g/L (CV <20%).
The dose-response of each hCG preparation (total, free [beta], nicked, and nicked free [beta]-hCG) was parallel to that of HhCG. Although hCG cross-reactivity was minimal, it is not clear whether such cross-reactivity was attributable to native hCG itself or to contamination of hCG preparations with HhCG.
In addition, stability studies using urine and serum samples from women with Down syndrome-unaffected pregnancies showed that HhCG was stable for 3 days at room temperature and 7 days in the refrigerator, as well as after two rapid freeze-thaw cycles.
(9) with the ELISA, we observed a similar increase in HhCG concentrations in Down syndrome patients.
In conclusion, we have developed an automated ICMA for the measurement of HhCG (ITA).
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