Discovery Of ApoM Gene Polymorphisms In Chronic Obstructive Pulmonary Disease And Establishment Of Detection Method

Objective:To detecte the SNPs of apoM gene in COPD patients and normal subjects,and established the typing methods of rs805264,rs707922 and rs707921.Methods:Six patients with COPD and normal subjects were tested for apoM gene polymorphisms by Roche 454 GS Junior high-throughput sequencer,then the base-quench probe techniqu was used to establish the corresponding typing method.ApoM gene rs805264,rs707922 and rs707921were detected in 252 COPD patients and 248 normal subjects by this method and the genotyping was confirmed by melting curves.The accuracy of the method was verified.Results:Total of 19 high-confidence mutations were detected in Roche 454,6 mutations among them were not reported in NCBI,but the mutation frequency was<20%.Four mutations occurred only in COPD patients,rs751064723 is locatedin the first exon of transcript 1,most of the rest are located in the apoM gene promoter region and intron region.The results of melting curve showed that the wild-type and homozygous mutants of rs805264,rs707922 and rs707921 showed melting valley at two different melting temperatures,and the results were consistent with the results of DNA sequencing(K=1,P 0.000).Conclusion:The discovery of apoM gene mutation sites in COPD patients provides a basis for further investigation of the relationship between apoM and COPD.The method for detecting single nucleotide polymorphisms is simple,economic and accurate,and it is suitable for a large number of apoM genotyping studies.