基于MLPA技术筛选帕金森病遗传相关基因及其与中医证型分布的关系和临床诊断价值

Screening of genetically related genes of Parkinson’s disease based on MLPA technology and their relationship with the distribution of traditional Chinese medicine(TCM)syndrome types and clinical diagnostic value

目的 探讨基于MLPA技术筛选帕金森病(PD)遗传相关基因及其与中医证型分布的关系和临床诊断价值。方法 选取2019年1月至2021年12月庐江县中医院确诊的106例PD患者作为研究对象,记为PD组,同时选取同期体检健康志愿者50名作为对照组,采用多重连接探针扩增(MLPA)技术检测,筛选出帕金森遗传相关基因,采用荧光定量(qRT)-PCR对基因mRNA表达水平进行检测。比较两组相关基因m RNA表达水平及相关基因与中医证型分布的关系,并绘制ROC曲线检验其对帕金森的临床诊断效能。结果 106例PD患者中36例(33.96%)PARKIN基因3号、4号外显子缺失突变,14例(13.21%)PARKIN基因2号外显子点突变,67例(63.21%)α-突触核蛋白(SNCA)基因1号外显子重复突变,其中15例(14.15%)患者未检出明确致病位点;PD患者不同的基因突变在证型的总体分布上,差异具有统计学意义(P<0.05);PD组外周静脉血PARKIN mRNA表达水平低于对照组、SNCA mRNA表达水平高于对照组,差异具有统计学意义(P<0.05);ROC曲线结果显示PARKIN mRNA、SNCA mRNA表达水平及其联合诊断曲线下面积(AUC)分别为0.727、0.859、0.905(P<0.05),敏感度分别为0.745、0.632、0.745,特异度分别为0.700、0.920、0.900。结论基于MLPA技术筛选出PD患者可能存在PARKIN基因与SNCA基因突变,且不同基因突变其中医证型分布不同,基因对应mRNA表达水平对PD具有一定的诊断价值。

Objective To explore the screening of genetic related genes of Parkinson ’s disease(PD)based on MLPA technique and their relationship and clinical diagnostic value with the distribution of TCM syndromes. Methods A total of 106 PD patients diagnosed in Lujiang County Hospital of Traditional Chinese Medicine from January 2019 to December 2021 were selected as the PD group,while 50 healthy volunteers from the same period medical examination were selected as the control group,the patients’ antecubital venous blood was examined by multiplex ligation probe amplification(MLPA),parkinsonism genetic related genes were screened,and the mRNA expression levels of genes were determined by fluorescence quantitative(q RT)-PCR. The mRNA expression levels of related genes were compared between the two groups,and the relationship between the related genes and TCM pattern distribution,and the ROC curve was plotted to test their clinical diagnostic efficacy for Parkinson’s. Results Thirty-six(33.96%)of 106 PD patients had deletion mutations in exons 3 and 4 of the PARKIN gene,14(13.21%)had point mutations in exon 2 of the PARKIN gene,and 67(63.21%)had point mutations α-Duplication mutation in exon 1 of the synuclein(SNCA)gene,of which 15(14.15%)patients had no clear pathogenic locus. Different gene mutations in PD patients showed statistical significance in the overall distribution of syndrome types(P<0.05). The PARKIN m RNA expression level in the peripheral venous blood in the PD group was lower than that in the control group,and the SNCA m RNA expression level was higher than that in the control group,with statistically significant differences(P<0.05).The ROC curve results showed that PARKIN m RNA,SNCA m RNA expression levels and their combined diagnostic area under the curve(AUC)were 0.727,0.859,0.905(P<0.05),with sensitivity of 0.745,0.632,0.745,and specificity of 0.700,0.920,0.900,respectively. Conclusion Patients with PD screened based on MLPA technique may have PARKIN gene mutations at SNCA gene,and different gene mutations have different distribution of TCM Patterns. The m RNA expression levels of the genes have a certain diagnostic value for PD.