Prothombin Time (PT) and Activated Partial Thromboplastin Time (APTT)

Question: Discuss about the Lab Report On Prothrombin Time (Pt) And Activated Partial Thromboplastin Time (Aptt)? Answer: Aim The primary aim of the study is to determine the prothombin time (PT) and activated partial thromboplastin time (APTT) from the supplied plasma samples. Introduction The entire mechanism of blood coagulation consists of various complex and dynamic interactions of platelets and blood plasma within the blood vessels(Polin, Fox and Abman, 2011). Blood coagulation plays an important contribution for haemostatic process and damage of protein wall results in activationof protein lipase enzyme and the final products become insoluble fibrin(Antovic and Blomback, 2010). The primary understanding of the coagulation pathway is to determine the prothombin and thromboplastin time results. For partial thromboplastin test there are mainly three categories are present such as intrinsic system, extrinsic system and common pathway. In this particular context, the modern coagulation diagnosis process has been effectively with prothombin time (PT) and activated partial thromboplastin time (aPTT)(Blomback and Antovic, 2009). The patients samples have been collected thoroughly and it helps in the detection of major disorders related to anticoagulation and clotting tim e. Hypothesis The effect of the changes of concentration of calcium chloride in the blood samples is directly proportional to the Prothrombin Time (PT) and the Activated Prothrombin Time (APTT). Methods PT test was performed on the plasma prepared from the collected blood samples and to perform the test effectively the initial arrangements were made. During the collection of blood,acid citrate dextrose is taken as anticoagulant(Tondre and Lebegue, 2010). Therefore, the separation of plasma and red cells are innovatively done and five healthy donors were taken for the blood samples and after that, plasma was dispensed into 5ml aliquots and frozen for the further process of the experiment(Oral Communication 3: Xeno Immunology - Non-Gal Antibodies and Coagulations (1), 2013). Then the plasma was incubated and CaCl2 is the added in regular interval and desired results has been collected (Oral Communication 5: Experimental Models and Non-Gal Antibodies and Coagulations (2), 2013).Statistical software has been utilized for thiscase to perform several statistical tests of the collected samples. Second groups of samples were collected for partial thromboplastin time deduction, which is also known as Activated Prothrombin Time test (APTT). Whereas, in Prothrombin time test, Calcium Chloride was added in order to determine the time taken for coagulation, in APTT, activated elements are added with the concentrations of calcium chloride. The samples are required top decalcified before so that they do not coagulate prematurely. The sample is plasma separated by centrifuging. The activated agents added are kaolin and cephalin. While kaolin works to activate the Factor XII and the cephalin serves as an alternative to the platelet phospholipids. In normal samples, the approximate time taken to clot is about 35 seconds. Results PT results The standard concentration of CaCl2 is 0.025M. In this particular context, different concentration of CaCl2 was added to each blood samples with different period and desired results have been collected. Time with CaCl2 Concentration Sample 1 (clotting Range) Sample 2 (clotting Range) Sample 3 (clotting Range) Sample 4 (clotting Range) Sample 5 (clotting Range) 1 min (0.025) 11 sec 9 sec 12 sec 10sec 10 sec 3 min (0.031) 10 sec 11 sec 11 sec 12 sec 11 sec 5 min (0.039) 9 sec 10 sec 10 sec 11 sec 8sec 7 min(0.089) 8 sec 12 sec 9 sec 9sec 7 sec 9 min (0.098) 7 sec 8 sec 6 sec 8 sec 6 sec APTT Results Time with CaCl2 concentration Sample 1 (clotting Range) Sample 2 (clotting Range) Sample 3 (clotting Range) Sample 4 (clotting Range) Sample 5 (clotting Range) 1 min (0.025) 35 sec 43 sec 34sec 38 sec 54 sec 3 min (0.346) 34 sec 41 sec 31 sec 36 sec 47 sec 5 min (0.426) 33 sec 38 sec 29 sec 33 sec 44 sec 7 min (0.589) 32 sec 36sec 28 sec 31sec 41 sec 9 min (1.255) 30 sec 32 sec 26 sec 24 sec 39 sec Parameter After 5 min After 8 min After 10 min PT (sec) Mean------------------------ Median--------------------- Min.max------------------- 10.291.36 11.78 9.69 -13.41 11.97 0.97 11.51 10.01-15.45 11.81 1.05 11.84 6.48-13.57 aPTT (sec) Mean SD------------ Median---------------- Min. Max------------- 46.697.76 48.26 36.46-69.91 54.03118.54 54.91 49.89-61.5 53.269.96 61.54 33.61-63.1 Significance: versus 5 min after collection P0.05 Effects of the variation in concentrations of calcium ions One of the important aspects of the tests is to determine the effects of the changes in the calcium chloride concentrations in the blood samples and in how they affected the Prothrombin time and the Activated Prothrombin Time of the samples. The hypothesis considered before the tests put down a directly proportional relation between the two. As the concentration of Calcium chloride was increased in the study of heparinized plasma, the APTT ratio indicated a likewise increase. The Calcium chloride concentrations recalcified the plasma-separated samples and served as a significant variable to manipulate and modify the sensitivity of the heparin aspect of APTT. An assay of 0.025 mol/L concentrations indicated increased APTT. Thus, the test results indicated a positive apprehension to the considered hypothesis. Conclusion The experiment provides a clear and concise idea regarding the PT and aPTT time for the given blood samples and from this, variation in coagulation of blood of different five healthy individuals are identified. It is evident from the study that increase in CaCl2 concentration results in reducing the PT an aPTT time of blood plasma. References Antovic, J. and Blomback, M. (2010).Essential guide to blood coagulation.Chichester, West Sussex, UK: Wiley-Blackwell. Blomback, M. and Antovic, J. (2009).Essential Guide to Blood Coagulation.Chichester: John Wiley Sons. Oral Communication 3: Xeno Immunology - Non-Gal Antibodies and Coagulations (1). (2013). Xenotransplantation, 20(5), pp.344-349. Oral Communication 5: Experimental Models and Non-Gal Antibodies and Coagulations (2). (2013). Xenotransplantation, 20(5), pp.356-360. Polin, R., Fox, W. and Abman, S. (2011). Fetal and neonatal physiology. Philadelphia: Elsevier/Saunders. Tondre, R. and Lebegue, C. (2010).Handbook of hematology research. New York: Nova Biomedical Books.