Clinical comparative study of HPV-DNA extraction by boiling method, magnetic bead method and centrifugal column method
应用煮沸法、磁珠法、离心柱法对HPV-DNA提取的临床对比研究

摘要

目的 探究应用煮沸法、磁珠法、离心柱法对HPV-DNA提取的临床对比价值。方法 选取2024年1月-2024年9月在我院体检中心、病房、门诊61例经检测HPV结果为阳性的宫颈脱落细胞样本，分别用于煮沸法、磁珠法、离心柱法进行DNA提取，并进行实时荧光定量PCR扩增，三种提取方法结果一致为阳性样本，比较其扩增后的循环阈值（Ct）差异和核酸提取效率。结果 三种提取方法经扩增后结果一致为阳性的样本为53例。对三组结果一致为阳性的53例样本结果进行统计学分析，ct值进行统计学分析无差异，DNA浓度和DNA纯度之间统计学有显著差异，煮沸法提取DNA浓度最高，离心柱法提取DNA纯度较好。结论 通过对比分析，煮沸法提取DNA浓度最高，但DNA纯度稍差，而离心柱法提取DNA纯度较好，但操作相对复杂，需要反复离心，不适合大批量提取，但其检出率较好，适合复查、样本量少的检测机构，磁珠法操作简单，适合大批量检测，但本实验采用的磁珠法CT值较其他两种方法学偏高，检出率偏低。

Abstract

Objective To explore the clinical comparative value of boiling method, magnetic bead method and centrifugal column method in the extraction of HPV-DNA.
Methods Sixty-one cervical exfoliated cell samples with positive HPV test results from the physical examination center, wards and outpatient department of our hospital from April 2021 to September 2024 were selected. DNA extraction was carried out by boiling method, magnetic bead method and centrifugal column method respectively, and real-time fluorescence quantitative PCR amplification was performed. The results of the three extraction methods were consistent and positive samples were identified. Compare the differences in the amplified circulation threshold (Ct) and the efficiency of nucleic acid extraction.
Results A total of 53 samples showed consistent positive results after amplification by the three extraction methods. Statistical analysis was conducted on the results of 53 samples with consistently positive results in the three groups. Statistical analysis of ct values showed no difference. There was a significant statistical difference between DNA concentration and DNA purity. The concentration of DNA extracted by boiling method was the highest, and the purity of DNA extracted by centrifugation column method was better.
Conclusion   Through comparative analysis, it is found that the boiling method has the highest DNA concentration, but the DNA purity is slightly lower. The centrifugation column method has better DNA purity, but the operation is relatively complex, requiring repeated centrifugation, and is not suitable for large-scale extraction. However, its detection rate is better, and it is suitable for re-examination and testing institutions with small sample volumes. The magnetic bead method is simple to operate and suitable for large-scale detection. However, the CT value of the magnetic bead method adopted in this experiment was higher than that of the other two methods, and the detection rate was lower.