Abstract
Authors reported the changes of F. XIII in human aortas with and/or without arteriosclerosis, in which we described some relation between arteriosclerosis and F. XIII. However, the measurement of F. XIII with Laurell method or radioisotope method is very troublesome and expensive. On the contrary, the single radial immunodiffusion method (Partigen method) is used easily and economically. The most troublesome problems which we encountered in Partigen method were the cloudy rings and/or too small rings caused by incorrect calculation. This study is to determine the optimal condition of F. XIII measurement using the Partigen method.
Methods and Materials
1) The fasting human plasmas were sampled from the antecubital vein. All samples were frozen at -20°C until use. Standard human plasma (from “Behringwerke”) was used as standard. In the Partigen plate, which had 1mm of thickness and 3mm of hole diameter, the amounts of anti-serum and/or the volume and concentration of sample plasma examined were altered. Namely, four different plates, in which 2.0%, 1.0%, 0.5% and 0.36% of anti-serum were included as the final concentrations. Preparing three different dilutions (two-fold, four-fold, and eight-fold) of plasma, 15μl, 30μl, 45μl, and 60μl, of each of the samples were injected into each of the holes. After 48hrs at room temperature, each of the plates were stained with tannic acid overnight. The diameter of the rings which appeared arround the holes were calculated.
2) The levels of F. XIII in several diseases were detected by the modified method mentioned above.
Resuts
a) Measurement of Sub-S
2.0% to 1.0% of anti-serum and 15μl to 30μl of sample with two-fold dilution were the most appropriate condition for the measurement of Sub-S.
b) Measurement of Sub-A
0.5% to 0.36% of anti-serum and 30μl to 60μl of sample with two-fold dilution were the most appropriate.
c) Correlation between Sub-A and Sub-S
The correlation of plasma F. XIII between Sub-A and Sub-S was r=0.381 (p<0.05). In one F. XIII dificient plasma, however, Sub-S was in the normal level, whereas Sub-A was not detected in this method.
d) The levels of F. XIII in several diseases
Normal range of our plasmas was in the 80% (standard as 100%). In the chronic liver disease, the level of F. XIII was lower, while the higher levels of F. XIII were obtained in hypertention, hyperlipidemia, diabetes mellitus, and collagen disease.
Conclusion
The optimal conditions for the measurement of plasma F. XIII were different in Sub-A and Sub-S. In our modified method, the reaction ring with the moderate diameter were obtained clearly. There was a clear correlation between Sub-A and Sub-S. In F. XIII deficient plasma, the fraction of Sub-A seemed to be damaged. The levels of plasma F. XIII in diseases should be continued to be studied to detect its variability and pathogenecity.
