Discussion Of The Role Of HLA-A MRNA In Host Peripheral Blood Lymphocytes In Actue Early Rejection After Renal Transplantation

Objects1. To develop a one-step real-time florescent quantitative RT-PCR (FQ-RT-PCR) method detecting human leukocyte antigen( HLA-A) mRNA of host peripheral blood T lymphocyte(PBL) before and after renal transplantation.Approaching the relationship among the HLA-A mRNA expression levels , the early actue rejection and the host immune state.2. Comparing with the indexes of HLA-A mRNA , the ratio of CD4/CD8, the blood Cr, BUN,the number of lymphocytes, evaluating the value of HLA-A mRNA in PBL of patients with renal transplantation as a early acute rejection index.MethodsAccording to HLA-A gene sequence announced in GenBank, the special primers and florescent probes targeting for the conservation districts in exon 4, 5 of HLA-A were synthesized and screened, glucose6-phosphate dehydrogenase (G6PDH) as inner reference; using Taqman probe, amplificated HLA-A mRNA and G6PDH mRNA by FQ-PCR. Getting the threshold cycle (Ct) of the amplification of 35 patients after renal transplantation in different time of half a year and 60 nomal samples, analyzed by REST software to get the relative expression of HLA-A mRNA. Considering the mean value(mV) which greater than or equal to normal value as the positive...