Preparation Of HLA Oligonucleotide Typing Microarry And Its Preliminary Use

Objective To establish HLA oligonucletide typing microarray and conduct someresearch work related to hereditary susceptibility of diabetic nephropathy by usingthis microarray platform. Contents The HLA typing is of great use in manyaspects including establishment of bone marrow stem cell bank, clarification ofmechanisms underlying of disease susceptivity and transplantation of organs andtissues. There are many polymorphism sites leading to the extreme difficulty.Serological method are usually adopted in clinical work to study HLA classâ… ,â…¡genes typing. However, The methods has shown several drawbacks. The majoyone is the antibody used are usually polyclonal and sometimes can cross reactnospecifically with other antigens in the blood, resulting in wrong conclusion. Analternative method for HLA typing is polymerase chain reaction withsequence-specific primers (PCR-SSP), which has already been employed in someexisting HLA typing kits. However, this method is never an ideal one since byusing this strategy, a number of electrophoreses have to be performed, whichconstrains its usefulness in clinical work. Since the 1990s, the microarraytechnology has undergone a great development. As the extending of dot blothybridization technology, microarray technology has been shown the great advantagein HLA typing because it is rather rapid and accurate making it very suitable for highthrough put screening of genes. We established HLA oligonucletide typingmicroarray and then conducted some research work related to hereditarysusceptibility of diabetic nephropathy by using this microarray platform. MethodsIn the present study, we have established a method to prepare oligonucleotide chipsby using 3-Aminopropyltriethoxysilane as the trip surface treatment reagent. Basedon the correlation of several hereditary diseas, e.g. Ankylosing Spondylitis andInfancy Diabetes, to HLA polymorphism. We designed 213 sequences specificoligonucleotide typing probes with medium distinguishability, most of which areconsist with the published sequences by the 13th HLA workshop. We haveconducted some research work related to hereditary susceptibility of diabetic nephropathy by using this rnicroarray platform. Results HLA classâ… ,â…¡alleleswere successfully typed in 30 clinical samples. This probe system is able to discern598 HLA classâ… alleles and 511 HLA classâ…¡alleles and proved that the DQA0501,DQB05 are highly correlated to diabetic nephropathy. Conclusion More ever, wehave proved that this test facility can be used not only to satisfied the need of geneticmatching but also to analyse the heredity factors of some autoimmune diseasescorrelation to HLA.