| 主权项 |
1. A method for determining the amount of fetal hemoglobin in an individual with sickle cell anemia undergoing hydroxyurea treatment, comprising:
a) providing a first sample from the individual comprising red blood cells and a second sample from the individual comprising red blood cells; b) adding the first sample to a buffer solution to form a first mixture, the buffer solution comprising an acidic buffer with a pH in a range of about 3.1 to about 3.5; c) incubating the first mixture for a sufficient time; d) subjecting the first mixture to centrifugation sufficient to form a solid deposit and a first supernatant; e) determining an optimum wavelength to measure absorbance of the first supernatant; f) determining an absorbance values of the first supernatant at the optimum wavelength, and determining the amount of sickle hemoglobin from a standard reference curve; g) adding the second sample to deionized water; h) incubating the second mixture for a sufficient time; i) subjecting the second mixture to centrifugation sufficient to form a solid deposit and a second supernatant; j) determining an optimum wavelength to measure absorbance of the second supernatant; k) determining an absorbance values of the second supernatant at the optimum wavelength, and determining the total amount hemoglobin from a standard reference curve; l) determining a relative amount of Fetal hemoglobin in the sample by subtracting the amount of sickle hemoglobin from the total hemoglobin. |
