A set of 15 proteins found in urine can distinguish healthy
individuals from those who have coronary artery disease (CAD), a new
study has found.
Due to the ease of obtaining samples, urinary protein analysis is
emerging as a powerful tool to detect and monitor disease. Anna
Dominiczak and colleagues tested whether urine could provide useful
biomarkers for coronary disease, one of the leading worldwide killers.
They analyzed samples from 88 CAD patients and 282 controls and found
a 15 protein "signature" indicative of disease. Several of the protein
fragments were collagens, which are components of arterial walls.
The researchers next examined how predictive their protein panel was
and found it could identify the presence of CAD 83% of the time. The
panel had a sensitivity of over 98%, which means the test produced
almost no false positives and thus inaccuracies are primarily
misdiagnosing CAD individuals as healthy. The researchers also
observed that the protein signatures of CAD individuals became more
normal after exercise, suggesting these biomarkers can be used to both
help diagnose CAD and monitor the progress of treatment
More bio-med news & videos
Portal to share biological information-data between people [Only registered users see links. ]
Pyrogallol red-molybdate(VI) method in microtiter plate techniques used as screening procedure for microalbuminuria determination PR reagent : 5.9 g of succinic acid and 0.5 g of sodium benzoate in 900 ml of water and mixing with 40 ml (24 mg/L correspondent to 0.06 mM) of dye solution (60 mg [1.5 mM] of Pyrogallol Red in 100 ml of methanol). Four milliliters of 2.4 g/L disodium molybdate and 8 ml of 35 g/L sodium oxalate (final concentration 2 mM for to reduce interference from aminoglycoside antibiotics can produce falsely increased urinary protein values) are added; the mixture is adjusted to pH 2.5 with 0.5 M HCl solution, dissolve 25 mg/L (0.1 mM) sodium dodecyl sulfate (SDS) as surfactant and brought to a volume of 1 L with deionized water. For to decrease interference by aminoglycoside eliminate addition of SDS and reduce to 20 ml (0.03 mM) dye solution. In consideration of SDS as interference value for patients treated with aminoglycoside antibiotics and its use to increase detectability of the γ-globulin, is need to realize two reagents in which : a) with specificity only against albumin molecule and, b) with specificity against albumin and γ-globulin proteins (total proteinuria). In the first reagent were omitted surfactant (SDS) while in the second 25 mg/L of SDS is added for simultaneous detected the greater urine proteins. From the difference of two values is possible to determinate γ-globulin proteins present in the sample.
BSA standard : consist of bovine serum albumin in concentrations of 0, 6.0, 12.5 25.0, 50.0, and 100 mg/L in distilled water or in 150 mM isotonic saline (NaCl) containing 1 g/L NaN3.
Urine samples are mixed by hand, then centrifuged at 4500-6000 rpm for 2 min. before being stored at -20 °C no more than three days before assay. Dispense in MTP, 25 µl of urine (duplicate), standards series (quadruplicate), urine-control (duplicate) and then quickly add in all wells 200 µl of pyrogallol red reagent without SDS and the plate allowed to stand at room temperature for 30 min., and Abs at 578 - 630 nm is measured.
Expected values/Reference range : 24-Hour Urinary Protein: 28 - 141 mg/day, Random Urine : <10 mg/dl