The Gene Detection Of CD36 Deficiency And Its Clinical Significance

Besides thrombopenia essentialis, many diseases have a symptom of thrombopenia. There are some serious consequences about thrombopenia in clinical. In recent years, with the development of blood transfusion, machine picks blood platelet transfusion has become one of the important methods to treat thrombocytopenia. However, with the increasement of infusion number, some chronic,long-term infusion patients appeared refractoriness to platelet transfusion(PTR). This problem is troublesome, causing blood transfusion world attention. The reasons of PTR is divided into the nonimmune factor and immune factors. platelet membrane antigen CD36 deficiency involved in the development of refractoriness to HLA and HPA-matched platelet transfusions. This phenomenon has become hot issue. The rate of CD36 defeciency, CD36 gene mutations and the PTR due to CD36 have been reported in abroad. These problems have not been reported in China. It is a blank field. We detect these problems and fill the blank in China. We hope that the detection of CD36 antigen can become a routine examined method. This can reduce platelet leosimendan-treated complications.Methods:300 healthy blood donors and 141 patients with thrombopenia were collected from The department of clinical laboratory, China-Japan union hospital, Jilin University from March 2009 to September 2010. The forty of these patients have more than twice infusion. Methods:①Genomic DNA were extracted from blood cells by Brother Biotech Blood DNA Kit.②The CD36 gene mutation was detected by a sequence-specific primers PCR(PCR-SSP) and positive cases with the mutation were checked by sequencing.③Analysis and compare the rate of CD36 deficiency between healthy and thrombopenia patients.④Study the platelet antibody with solid-phase Coombs Test. Guess the relationship between PTR and CD36 deficiency.Results:Among 300 healthy blood donors are found in 19 existing four kinds of gene mutations. These gene mutations conclude the C→T substitution at 267 site,deletion GC at 329 site,the insertion T at 560 site,T→G substitution at 1079 site. Among 141 patients are found in 9 existing the same four kinds of gene mutations. Among 300 healthy blood donors, there are three donors existed the gene mutation of C for T substitution at 267 site accounted 15.8% in mutated; there are ten donors existed the gene mutation of deletion GC at 329 site accounted 52.6% in mutated; there are one donor existed the gene mutation of the insertion T at 560 site accounted 5.3% in mutated; there are five donors exested the gene mutation of the T for G substitution at 1079 site accounted 26.3% in mutated. Among 141patients, there are two patients existed the gene mutation of C for T substitution at 267 site accounted 22.2% in mutated; there are three patients existed the gene mutation of deletion GC at 329 site accounted 33.3% in mutated; there are three patients existed the gene mutation of the insertion T at nt560 accounted 33.3% in mutated; there are one patient existed the gene mutation of the T for G substitution at nt1079 accounted 11.1% in mutated. That is to say, nine patients may be typeⅡCD 36 deficiency among 141patients.The two groups of the healthy blood donors group and the patients group are detected out the same four gene mutations. Compare the constituent ratio of both the healthy blood donors group and the patients group with Chi square test. The result of comparison is P>0.05.There is no different in the constituent ratio of the two groups.141 patients were detected by platelet antibody(HLA and HPA) kit. There are 101 patients accepted platelet transfusion once. Among these patients are found 4 people CD36 deficiency. There are 40 patients accepted platelet transfusion more twice. according to the judgement standard of PTR, there are 25 patients have good results,15 patients occur PTR. Among 25 patients are no found CD36 deficiency and platelet antibody detection negative. Among 15 patients, there are 7 patients have no immune factors; 3 patients existed HLA/HPA positive; 2 patients existed HLA/HPA positive and CD36 deficiency; 3 patients only existed CD36 deficiency. According to this result, we have reason to doubt that the CD36 deficiency can cause PTR, but we have other evidence and a large number of experiment.Conclusion:1, CD36 deficiency rate is 6.3%.2, There are 4 gene mutations can cause CD36 deficiency:These gene mutations conclude the C→T substitution at 267 site,deletion GC at 329 site,the insertion T at 560 site,T→G substitution at 1079 site.3, gene mutation of 267 site may not be the most important reason of CD36 deficiency in Chinese, while the gene mutation of deletion GC at 329 site seems more important for CD36 deficiency in Chinese. The gene mutations of insertion T at 560 site and T for G substitution at 1079 site exist in our detected samples. The two gene mutations are rare been reported in all kinds of human race which may be exist some different in human races in CD36 deficiency.4, Thrombocytopenia can not influence expression of CD36. thrombocytopenia have no direct relationship with CD36 deficiency. CD36 deficiency cause by gene mutations. CD36 deficiency influence the effect of the platelet transfusion.5, CD36 deficiency may have relationship with PTR. The detection of CD36 deficiency will be do before the platelet transfusion to matched platelet transfusion. It can improve the effect of transfusion.