Detecting proteinuria in urine is an important biomarker in medicine. Increased levels of proteins may indicate kidney disease. Two samples of urine were delivered marked as ‘patient’ & ‘healthy. The patient complained about fever and tiredness after endurance, and stated that blood was found in the urine.
To assess the concentration of proteins, to provide scientific reasoning based on data obtained. Comparison of samples by double-tailed T-Test to reveal the significance of results. To assess the patient’s case.
It is expected to assess the concentration of proteins in both samples. To assess the significance of the results.
Materials and methods
A quantitative Bradford assay was used for the estimation of proteins. Thus, a dye known as Coomassie blue provides a colorimetric time-tested change from red to blue when it binds to proteins under acidic conditions. Spectrometry was used to assess absorbance. Known concentration of Bovine Serum Albumin (BSA) was used as a reference point to assess unknown protein concentrations in both samples. Standard dilutions of Bradford and BSA were prepared.Absorbing readings were normalized with blank. Spectrometer set to 595 nm wavelength. All spectrometry observation was triplicate repeated, all results were recorded. Incubation time at room temperature 5min.
Concentrations were deduced based on the linear equation after setting up the intercept. y=mx where y=absorbance, x=concentration, m=slop. According to Chart No. 1 calibration, standard curve was plotted based on provided concentration of BSA. Unknow protein concentrations were deduced from rearranged linear equations displayed in Chart No. 1. Results with standard deviation are presented in Table No.1 T-test was used as a statistical tool. Significance was marked on Chart No. 2. Urine protein levels have significance resulted for P. Each group n=3 two-tailed t-test. Obtain data are presented as mean ± standard deviation. t(4)=(-34.87), P<0.00001. Therefore, P****.
Chart No. 1
|Tested samples Protein concentration in urine (mg/liter ± standard deviation) Healthy sample 1.4003 ± 0.007 Patient sample 1.5823 ± 0.005 Table No.1||Chart No. 2|
Discussion and conclusion
All objectives of the analysis were reached. According to Aitekenov, (2020), used methodology is similar. Two points on the calibration curve do not match the trend line. There is a probability that the researcher used incorrect volume in those two calibration BSA samples. Comparison of healthy sample versus patient sample reveals that level of protein is abnormal to patient. Statistical verification by a two-tailed t-test revealed the significance factor of obtaining data. Blood in urine and other patient symptoms lead to the conclusion of glomerulonephritis. Additional screenings should proceed to find the cause of the patient’s condition. Accuracy & efficiency may be improved with the adoption of different screening methods according to Schleicher, (1978).
- Aitekenov, S. et al. (2020), Detection and quantification of proteins in human urine. Talanta, p.121718. [online] Available at: https://www.sciencedirect.com/science/article/pii/S0039914020310092?via%3Dihub [Accessed 29 Nov. 2021].
- Schleicher, E. and Wieland, O.H. (1978), Evaluation of the Bradford method for protein determination in body fluids. Journal of Clinical Chemistry and Clinical Biochemistry. Zeitschrift Fur Klinische Chemie Und Klinische Biochemie, [online] 16(9), pp.533–534. Available at: https://pubmed.ncbi.nlm.nih.gov/712344/ [Accessed 29 Nov. 2021].