Improved one-step micro-lymphocytotoxicity cross-over trial.docVIP

 PAGE \* MERGEFORMAT 5 Improved one-step micro-lymphocytotoxicity cross-over trial Lymphocytotoxicity cross-over trial, also known as complement-dependent cytotoxicity test (CDC). It is to check whether the body organ transplant recipients for the donor or donors for the recipient’s HLA antibodies in organ transplant must be carried out before the CDC test. If detected by the serum against the donor’s HLA antibodies, is generally considered taboo with transplant recipients occur after the hyperacute rejection, leading to graft failure. As the pilot implementation of an organ transplant operation in the interim before the match of the test results of the report requested in a short time, so we made an improvement of the technology compared with the traditional method, you receive satisfactory results. Are reported as follows: One, materials and methods 1. Was seized object: specimens from the past 10 months in our hospital inpatient and outpatient 280 cases of renal transplantation to be done to be medical consultation and other organ transplants to be patients attending who do not adopt 2 ml blood anticoagulant under test. 2. Reagent: Immunization beads, fluorescent dye liquor OL provided by the United States, anti-lymphocyte antibody reagents all purchased from Wuhan Institute of Biological Products. 3. Methods: (1) Improvement step: use of immunomagnetic beads 3 ~ 5 μ l isolated lymphocytes, in the Terasaki plate sterile mineral oil per well by adding 5 μ l, each row of the first one-hole plus 1 μ l saline so negative contrast, the first two holes plus 1 μ l of anti-lymphocyte antibody-wide to do positive control, 3 to 6 holes of various increases in serum by 1 μ l (ie repeat the 4-hole tests), each hole for each category of donor lymphocytes in 1 μ l (cell concentration 2 000 / μ l), each well complement each plus 1 μ l, home 22 ~ 25 ℃ room temperature for about 45 min reaction, each well add fluorescent dye liquor 5 μ l, the cells were observe