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Prostatic antigens (Psa) are tumor markers that help monitor the state of the prostate, as well as determine the progress in case of cancer in that area. These markers have two fractions; total and free. These tests are performed with a blood sample in a sterile tube without additives, separating the sample to later obtain the serum.
In human serum the psa is found in several forms, it can be free or forming a stable complex with the ACT. The total psa is the sum of those two fractions (Psa-L + Act). It is important to note that when making a PSA report, this value cannot exceed that of the PsaT.
Today we will be studying the processing to determine the concentration of free Psa in human serum by Elisa's method
- The first step is to temper the reagents to be used, in this case I will work with the Vitrotest frame. These should be at room temperature for approximately 30 minutes to 1 hour. Later I check that my box has all the necessary reagents. I prepare the standards, controls and washing solutions to create my curve from the beginning.
- Once I have all my materials prepared and having obtained my patient's serum, we proceed to read the technique. Despite having made this curve many times it is always important to read the techniques, since the quantities may have varied between batches of reagents.
The summarized technique always placed them in the boxes to have at hand and not waste time, it is already crucial to be precise and exact when advancing in our steps. In this case, the curve is created with 6 standards plus the control well, so in total I will use 8 wells, counting the patient.
Once the grid is ready, the quantities of standard and AntiPsa established by the technique are poured. For the formation of the antigen-antibody complex. The antigen binds to the solid plate that the well has.
I will use the water bath at a temperature of 37 degrees to carry out the incubations. The first is exactly 30 minutes, the time I've been on a stopwatch. It is extremely important to respect the times as indicated by the technique to obtain correct results.
Once the time has passed, the first wash is carried out. With approximately 300 moors of prepared water and 5 washes. After drying the wells very well, the conjugate or enzyme is added, left to incubate at temperature for another 30 minutes.
- After that time, the washing is carried out again x 5 times, it is dried again and the TMB or chromogen is added, this solution allows the complexes formed to be colored.
— It is incubated at room temperature for 15 minutes, then the stop solution is added and the reaction is stopped. This curve is directly proportional to the intensity of color that is obtained, that is, the more intense the color, the higher the concentration of the antigen.
- After performing all the technique, we proceed to read the absorbances. In this case I will use a Stat Fax that allows me to read the absorbances and calculate the concentrations directly with the information that I provide you from the known concentrations of the standards.
It is important to note that correct results are not only obtained by a processing game, but also by a good sample collection and the previous preparation of the patient. In this case the patient must be fasting, have sexual abstinence, not having used bicycles or riding a horse and not having had a prostate echo in approximately 3 to 5 days.
Not least, these techniques should only be performed by specialized health personnel.
- Traductor DeepL
- Contenido de mi Autoria.
- Fotografía tomadas con mi iPhone 12 pro
-Medios de edición: Canva
