粪便中脱落细胞基因检测在炎症性肠病诊断中的临床价值

Clinical value of gene detection of exfoliated cells in feces in the diagnosis of inflammatory bowel disease

目的探索粪便中脱落细胞基因检测检测在炎症性肠病诊断中的临床价值。方法选取2020年6月至2021年6月期间在深圳市第二人民医院收治的30例健康人群作为对照组,70例炎症性肠病患者作为观察组,收集所有受检者粪便样本后,进行粪便中DNA的提取和荧光定量PCR检测,经ROC曲线分析SDC2甲基化水平、内参基因ACTB预测效能,并用Spearman法分析内参基因ACTB表达与临床病理特征关系。结果两组SDC2甲基化检测Ct值、阳性率比较,差异无统计学意义(P>0.05),观察组ACTB的阳性率高于对照组,ACTB Ct值低于对照组,差异有统计学意义(P<0.05)。经ROC曲线分析,内参基因ACTB、SDC2甲基化Ct值预测炎症性肠病的AUC分别为0.918、0.473;不同性别、年龄段患者、内参基因ACTB的Ct值比较,差异无统计学意义(P>0.05),不同病程、病变位置患者内参基因ACTB的Ct值比较,差异有统计学意义(P<0.05)。经Spearman法分析,内参基因ACTB与病程、病变位置呈正相关性(P<0.05)。结论粪便中脱落细胞内参基因ACTB表达水平与肠炎可能有关系,SDC2甲基化与肠炎无关系。

Objective To explore the clinical value of detection of exfoliated cells in feces in the diagnosis of inflammatory bowel disease.Methods Seventy patients with inflammatory bowel disease admitted to Shenzhen Second People’s Hospital from June 2020 to June 2021 were selected as the observation group,and 30 healthy people were selected as the control group.The stool samples were collected for DNA extraction and fluorescence quantitative PCR(qPCR)detection.The ROC curve was used to analyze the methylation of SDC2 and internal reference gene ACTB was used to predict assay performance.Spearman method was used to analyze the relationship between the expression of internal reference gene ACTB and clinicopathological characteristics.Results There was no significant difference in the Ct value and positive rate of SDC2 methylation detection between the two groups(P>0.05).The positive rate of ACTB in the observation group was higher than that in the control group,and the ACTB Ct value was lower than that in the control group.The difference was statistically significant(P<0.05).Based on ROC curve analysis,the methylation Ct values of internal reference genes ACTB and SDC2 predicted the AUC of inflammatory bowel disease to be 0.918 and0.473,respectively.There was no statistically significant difference in the Ct values of patients with different genders,ages,and internal reference gene ACTB(P>0.05).The comparison of the Ct values of internal reference gene ACTB in patients with different course of disease and lesion location showed statistically significant differences(P<0.05).According to Spearman.s analysis,the internal reference gene ACTB was positively correlated with the course of the disease and the location of the disease(P<0.05).Conclusion The expression level of the reference gene ACTB in fecal exfoliated cells may be related to enteritis,while SDC2 methylation has no relationship with enteritis.