目的:探讨荧光定量PCR法与涂片抗酸染色法在活动性肺结核早期诊断中的应用价值,评估两种方法在敏感性、特异性及准确性方面的表现,为临床诊断提供科学依据。方法:研究选取2021年1月至2022年1月期间我院收治的100名疑似活动性肺结核患者...目的:探讨荧光定量PCR法与涂片抗酸染色法在活动性肺结核早期诊断中的应用价值,评估两种方法在敏感性、特异性及准确性方面的表现,为临床诊断提供科学依据。方法:研究选取2021年1月至2022年1月期间我院收治的100名疑似活动性肺结核患者作为研究对象,根据检测方法分为实验组和对照组,实验组采用荧光定量PCR法进行检测,对照组则采用涂片抗酸染色法进行诊断。检测前均采集患者的痰液样本并进行标准化处理。记录两组的阳性检出率、敏感性、特异性及结果一致性。结果:荧光定量PCR法的敏感性为90.48%,特异性为93.75%;涂片抗酸染色法的敏感性为66.67%,特异性为87.50%。荧光定量PCR法在检出真实阳性病例方面表现更优,假阴性率更低,同时在排除非病原体感染的能力上表现更高,假阳性率较低。两种方法对比表明,荧光定量PCR法在敏感性和特异性方面均优于涂片抗酸染色法,具有更高的诊断准确性和临床应用价值(P Objective: To explore the application value of fluorescence quantitative polymerase chain reaction (PCR) and Acid-Fast Smear Staining in the early diagnosis of active pulmonary tuberculosis, to assess the performance of the two methods in terms of sensitivity, specificity and accuracy, and to provide scientific basis for clinical diagnosis. Methods: 100 patients with suspected active tuberculosis admitted to our hospital between January 2021 and January 2022 were selected as the study subjects, and were divided into experimental and control groups according to the detection methods, with the experimental group using the fluorescence quantitative PCR for the detection, and the control group using the smear antacid staining method for the diagnosis. Sputum samples were collected and standardized before testing. The positive detection rate, sensitivity, specificity and consistency of the results of the two groups were recorded. Results: The sensitivity of the fluorescence quantitative PCR was 90.48% and the specificity was 93.75%;the sensitivity of the Acid-Fast Smear Staining was 66.67% and the specificity was 87.50%. The fluorescence quantitative PCR performed better in detecting true-positive cases with a lower false-negative rate, as well as higher in the ability to exclude non-pathogenic infections with a lower false-positive rate. Comparison of the two methods showed that the fluorescent quantitative PCR was superior to the Acid-Fast Smear Staining in terms of sensitivity and specificity, with higher diagnostic accuracy and clinical application value (P < 0.05). Conclusion: Although the Acid-Fast Smear Staining is easy to operate, it has limited sensitivity and accuracy and is not suitable for early screening application. It is recommended that the fluorescence quantitative PCR be used as the first choice for the early diagnosis of active pulmonary tuberculosis to improve the diagnostic efficiency and provide a precise basis for the early treatment of patients.展开更多
目的:使用高通量测序法辅助开发RAS相关区域家族1A(Ras-Association Domain Family 1A,RASSF1A)基因甲基化定性检测试剂盒(荧光PCR法)。方法:提取37例肺癌血浆样本DNA进行亚硫酸氢盐修饰转化,使用高通量测序方法检测样本的亚硫酸氢盐转...目的:使用高通量测序法辅助开发RAS相关区域家族1A(Ras-Association Domain Family 1A,RASSF1A)基因甲基化定性检测试剂盒(荧光PCR法)。方法:提取37例肺癌血浆样本DNA进行亚硫酸氢盐修饰转化,使用高通量测序方法检测样本的亚硫酸氢盐转化效率和RASSF1A基因CpG位点的甲基化水平,之后使用荧光PCR法检测样本RASSF1A基因的甲基化状态。结果:高通量测序显示样本DNA的平均转化效率为99.47%;根据CpG位点甲基化水平的双向聚类分析结果,筛选出第26~30号CpG位点作为荧光PCR法的检测位点,以甲基化水平5%作为阈值线,ΔCt Cut-off值为9,得出两种检测方法对37例样本的测定结果一致。结论:本研究借助高通量测序技术成功构建基于荧光PCR法的RASSF1A基因甲基化定性检测方法,操作简便、成本低、效率高,适用于临床RASSF1A基因甲基化状态常规化检测。展开更多
文摘目的:探讨荧光定量PCR法与涂片抗酸染色法在活动性肺结核早期诊断中的应用价值,评估两种方法在敏感性、特异性及准确性方面的表现,为临床诊断提供科学依据。方法:研究选取2021年1月至2022年1月期间我院收治的100名疑似活动性肺结核患者作为研究对象,根据检测方法分为实验组和对照组,实验组采用荧光定量PCR法进行检测,对照组则采用涂片抗酸染色法进行诊断。检测前均采集患者的痰液样本并进行标准化处理。记录两组的阳性检出率、敏感性、特异性及结果一致性。结果:荧光定量PCR法的敏感性为90.48%,特异性为93.75%;涂片抗酸染色法的敏感性为66.67%,特异性为87.50%。荧光定量PCR法在检出真实阳性病例方面表现更优,假阴性率更低,同时在排除非病原体感染的能力上表现更高,假阳性率较低。两种方法对比表明,荧光定量PCR法在敏感性和特异性方面均优于涂片抗酸染色法,具有更高的诊断准确性和临床应用价值(P Objective: To explore the application value of fluorescence quantitative polymerase chain reaction (PCR) and Acid-Fast Smear Staining in the early diagnosis of active pulmonary tuberculosis, to assess the performance of the two methods in terms of sensitivity, specificity and accuracy, and to provide scientific basis for clinical diagnosis. Methods: 100 patients with suspected active tuberculosis admitted to our hospital between January 2021 and January 2022 were selected as the study subjects, and were divided into experimental and control groups according to the detection methods, with the experimental group using the fluorescence quantitative PCR for the detection, and the control group using the smear antacid staining method for the diagnosis. Sputum samples were collected and standardized before testing. The positive detection rate, sensitivity, specificity and consistency of the results of the two groups were recorded. Results: The sensitivity of the fluorescence quantitative PCR was 90.48% and the specificity was 93.75%;the sensitivity of the Acid-Fast Smear Staining was 66.67% and the specificity was 87.50%. The fluorescence quantitative PCR performed better in detecting true-positive cases with a lower false-negative rate, as well as higher in the ability to exclude non-pathogenic infections with a lower false-positive rate. Comparison of the two methods showed that the fluorescent quantitative PCR was superior to the Acid-Fast Smear Staining in terms of sensitivity and specificity, with higher diagnostic accuracy and clinical application value (P < 0.05). Conclusion: Although the Acid-Fast Smear Staining is easy to operate, it has limited sensitivity and accuracy and is not suitable for early screening application. It is recommended that the fluorescence quantitative PCR be used as the first choice for the early diagnosis of active pulmonary tuberculosis to improve the diagnostic efficiency and provide a precise basis for the early treatment of patients.
文摘目的:使用高通量测序法辅助开发RAS相关区域家族1A(Ras-Association Domain Family 1A,RASSF1A)基因甲基化定性检测试剂盒(荧光PCR法)。方法:提取37例肺癌血浆样本DNA进行亚硫酸氢盐修饰转化,使用高通量测序方法检测样本的亚硫酸氢盐转化效率和RASSF1A基因CpG位点的甲基化水平,之后使用荧光PCR法检测样本RASSF1A基因的甲基化状态。结果:高通量测序显示样本DNA的平均转化效率为99.47%;根据CpG位点甲基化水平的双向聚类分析结果,筛选出第26~30号CpG位点作为荧光PCR法的检测位点,以甲基化水平5%作为阈值线,ΔCt Cut-off值为9,得出两种检测方法对37例样本的测定结果一致。结论:本研究借助高通量测序技术成功构建基于荧光PCR法的RASSF1A基因甲基化定性检测方法,操作简便、成本低、效率高,适用于临床RASSF1A基因甲基化状态常规化检测。
