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Department of Internal Medicine I, University Hospital Dijkzigt, Dr. Molewaterplein 40, 3015 GD Rotterdam, The Netherlands.
a Author for correspondence. Fax 31 10 4634531; e-mail deinum{at}inw1.azr.nl
Background: Our renin IRMA overestimated renin in plasmas with high prorenin-to-renin ratios. We suspected that the overestimation of renin was caused less by cross-reactivity of the renin-specific antibody with prorenin than by a conformational change of prorenin into an enzymatically active form during the assay.
Methods: Because the inactive form of prorenin converts slowly into an active form at low temperature, we raised the assay temperature from 22 °C to 37 °C, simultaneously shortening the incubation time from 24 to 6 h. The former IRMA was performed in <1 working day with these modifications.
Results: The comeasurement of prorenin as renin was eliminated. Reagents were stable at 37 °C, and the new and old IRMAs were comparable in terms of precision and accuracy. The functional lower limit of the assay (4 mU/L) was below the lower reference limit (9 mU/L). The modified IRMA agreed closely with the activities measured with an enzyme-kinetic assay. Results were not influenced by the plasma concentration of angiotensinogen. At normal angiotensinogen concentrations, the IRMA closely correlated with the classical enzyme-kinetic assay of plasma renin activity.
Conclusion: The modified IRMA, performed at 37 °C, avoids interference by prorenin while retaining the desirable analytical characteristics of the older IRMA and requiring less time.© 1999 American Association for Clinical Chemistry
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