Construction Of Gene Microarray For Detection Of Differentiated Thyroid Cancer Related Genes

Background:Thyroid cancer accounts for 1% of human malignant tumors,and is the most common malignant tumor of the endocrine system.The incidence of the disease is rising around the world,and has become the most rapidly growing human tumor compared with other malignant tumors.At present,ultrasound examination,fine needle aspiration cytology and serum thyroglobulin detection are common methods for the diagnosis of thyroid tumors,however,because of the existence of misdiagnosis and missed diagnosis,the establishment of more accurate and specific diagnostic methods has become the urgent problem to be solved.Mutations in BRAF,RAS and TERT promoter regions have an inestimable role in the pathogenesis of thyroid cancer due to abnormal cell proliferation,in recent years,with the application of molecular diagnostic techniques in the diagnosis and treatment of tumors,the detection of BRAF,RAS and TERT promoter mutations in thyroid cancer has been carried out in clinical practice.However,due to the false positive/negative results caused by the doctor's technical experience and inspection equipment,a new method of high sensitivity and specificity should be studied.Objective:To establish a new type of thyroid cancer related gene detection technology,improve the sensitivity and specificity of clinical testing and provide the basis for clinical diagnosis and treatment.Methods:The synthesis of citrate stabilized gold nanoparticles for the construction of gold nanoparticle labeled probe function;preparation of DNA microarray for the detection of target DNA;silver enhancement reaction to amplify the signal;colorimetricmicroarray DNA microarray scanner to obtain RLS signal;Image J extracts the value of the RLS signal and background signal value for data analysis.Results:When the chip sampling probe concentrations were: Probe 1T and 1S :30 ?M,Probe2 T,2S,3T,3S:5?M;hybrid buffer solution is 3×SSC,0.1%(w/v)SDS;functionalized gold nanoparticles buffer solution is 1× SSC,0.1%(w/v)SDS.The hybridization reaction was performed at 45 C for 2 hours,functionalized gold nanoparticles react at 30 C for 1 hours.More stable and specific results can be obtained.Conclusions:The application of DNA microarray combined with functionalized gold nanoparticles in the detection of thyroid cancer related genes is a highly sensitive and specific method,which has a good reference value for clinical diagnosis and treatment.