Detection of Tissue Factor Pathway Inhibitor-2 Gene Mutation and its Correlation with Coagulaton Parameters in Sudanese Pediatric Patients with Leukemia

Abstract

This is analytical case control study was carried out in radio isotope center Khartoum (RICK) to detect TFPI-2 gene mutation and its correlation with coagulation parameter in Sudanese pediatric patients with leukemia. Ninety-eight patients less than 15 years with mean age for all (9.03 ± 4.058) where diagnosed with leukemia of both gender 52 (53.1%) were males and 46 (46.9%) were females .The center administration and the relatives of the patients were informed about the study and agreement of participation was obtained. In additive ninety-seven healthy children, less than 15 years 53 (54.6%) were males and 44 (45.4%) were females with mean age for all (8.08 ± 4.033) selected as a control group. A structured questionnaire used to collect information about patient age, sex, and patient history of disease. Five ml of venous blood was collected, three ml in EDTA and 1.8 ml in trisodium citrate anticoagulant container from all patients and controls. Reverse transcription polymerase chain reaction followed by restriction-fragment length polymorphism (RFLP) analysis of the TFPI-2 was, carried out for all patients and controls samples. TFPI-2 gene was investigated in all samples, there were 14(14.3%) of the patients group have mutation, 10(71.4%) of them were homozygous and 4 (28.6%) were heterozygous. No participant among control group (0%) had TFPI-II gene mutation; there was significant association between patients, which have TFPI-2 gene mutation, and control group, P value 0.0258. Automated hematological analyzer (Sysmex KXN-21) used to measure platelet count for the patient and control group. Automated coagulation tests (PT, PTT, fibrinogen, protein C and D-dimer level) were measured using coagulation analyzer (sysmex CA 500) for all patient and control group. Statistical package for social science (SPSS) computer program was used for data analysis In patients group with TFPI-2 gene mutation the result of PT mean level was (17.05 ± 2.974). PTT mean level was (39.564 ± 6.809), fibrinogen mean level (2.759 ± 0.630), protein C mean level (0.845± 0.198) and platelet count mean (288.21 ± 155.797), while in the patients group without mutation the result of, PT mean was (17.689 ± 6.741), PTT mean was (40.555 ± 11.551), fibrinogen mean (2.659 ± 0.681), protein C mean level (0.822 ± 0.037) and platelet count mean was(289.67 ± 136.85). There were insignificant variations in coagulation tests parameter (platelet count, PT, INR, APTT, protein C and fibrinogen) between patients with TFPI-2 gene mutations and those without TFPI-2 gene mutations. There were only two patients had high D-dimer level which excludes disseminated intravascular coagulation.