[1]葛 敏,陈雁楠,吴长宇,等.基于拉曼光谱分析技术建立快速筛查高危型HPV感染的方法及临床应用研究[J].现代检验医学杂志,2026,41(02):10-15+36.[doi:10.3969/j.issn.1671-7414.2026.02.003]
　GE Min,CHEN Yannan,WU Changyu,et al.Establishment of a Rapid Screening Method for High-Risk HPV Infection Based on Raman Spectroscopy Analysis and Its Clinical Application Research[J].Journal of Modern Laboratory Medicine,2026,41(02):10-15+36.[doi:10.3969/j.issn.1671-7414.2026.02.003]
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基于拉曼光谱分析技术建立快速筛查高危型HPV感染的方法及临床应用研究()

《现代检验医学杂志》[ISSN:/CN:]

卷:
第41卷
期数:
2026年02期
页码:
10-15+36
栏目:
论著
出版日期:
2026-03-15

文章信息/Info

Title:
Establishment of a Rapid Screening Method for High-Risk HPV Infection Based on Raman Spectroscopy Analysis and Its Clinical Application Research
文章编号:
1671-7414（2026）02-010-07
作者:
葛 敏1，陈雁楠2，吴长宇2，耿志欣3，裴 兵1,3
1.扬州大学医学院，江苏扬州 225009;2.徐州医科大学医学影像学院，江苏徐州 221000;3.南京医科大学附属宿迁第一人民医院，江苏宿迁 223800
Author(s):
GE Min1，CHEN Yannan2，WU Changyu2，GENG Zhixin3，PEI Bing1,3
1.Yangzhou University Medical College, Jiangsu Yangzhou 225009, China;2.School of Medical Imaging, Xuzhou Medical University, Jiangsu Xuzhou 221000, China;3.the Affiliated Suqian First People’s Hospital of Nanjing Medical University, Jiangsu Suqian 223800, China
关键词:
高危型人乳头瘤病毒; 拉曼光谱分析技术; 快速筛查; 临床验证
分类号:
R373.9；R392-33
DOI:
10.3969/j.issn.1671-7414.2026.02.003
文献标志码:
A
摘要:
目的?探索基于拉曼光谱分析技术建立快速筛查高危型人乳头瘤病毒(HPV)感染的方法,并通过临床样本验证其可行性及有效性。方法选取南京医科大学附属宿迁第一人民医院门诊就诊的300例女性患者,根据HPVDNA检测结果分为HPV阴性对照组(n=100)、HPV16/18阳性组(n=100)和其他高危型HPV阳性组(n=100)。采用枸橼酸钠还原法合成金纳米颗粒,用4-巯基苯甲酸(4-MBA)、4-氨基硫代苯酚(4-ATP)和2-萘硫醇(2-NT)三种拉曼报告分子标记,再偶联针对HPV16L1蛋白、HPV18L1蛋白和通用高危型HPVE6/E7蛋白的单克隆抗体,构建三种拉曼活性探针。采用共聚焦拉曼光谱仪采集样本光谱,通过支持向量机(SVM)建立HPV分型模型。对临床标本进行检测并与传统HPVDNA检测方法进行比较及性能评价。结果三种拉曼活性探针光谱特性良好,特征峰无重叠。构建的SVM模型分类准确度为93.0%,对HPV16、HPV18及其他高危型HPV的分型准确度分别为95.2%、93.7%和91.4%。受试者工作特征(ROC)曲线分析显示,检测曲下面积(AUC)值分别为0.978、0.962和0.943。拉曼光谱检测方法的灵敏度、特异度、准确度分别为92.5%、94.3%、93.3%,Kappa系数为0.865。高危型HPV检测下限为0.1IU/ml,变异系数为4.3%;温度(4℃~37℃)稳定性测试无显著差异(P>0.05);干扰试验显示,血红蛋白和白蛋白信号偏差分别控制在8%和6%以内,白细胞介素-6(IL-6)和肿瘤坏死因子-α(TNF-α)信号偏差分别小于7%和6%。连续30天监测显示,探针拉曼信号强度衰减率为3.8%,稳定性良好。检测时间缩短至45.3±6.7min,成本降至每样本4元。结论基于拉曼光谱分析技术的高危HPV检测方法具有良好的灵敏度、特异度和分型能力,与传统方法相比,具有检测时间短、成本低、特异度高等优势,具备广阔的临床应用前景。
Abstract:
Objective To explore a rapid screening method for high-risk human papillomavirus (HPV) infection based on Raman spectroscopy analysis and validate its feasibility and effectiveness using clinical samples. Methods A total of 300 female outpa-tients from Suqian First People’s Hospital Affiliated to Nanjing Medical University were enrolled and divided into three groups according to HPV DNA test results: HPV-negative control group (n=100), HPV16/18-positive group (n=100), and another high-risk HPV-positive group (n=100). Gold nanoparticles were synthesized using the sodium citrate reduction method, labeled with three Raman reporter molecules: 4-mercaptobenzoic acid (4-MBA), 4-aminothiophenol (4-ATP), and 2-naphthyl mercaptan (2-NT), and then conjugated with monoclonal antibodies targeting HPV16 L1 protein, HPV18 L1 protein, and pan-high-risk HPV E6/E7 proteins to construct three Raman-active probes. Confocal Raman spectroscopy was used to collect sample spectra, and a support vector machine (SVM) was used to establish an HPV genotyping model. Clinical specimens were detected and compared and per-formance evaluated with conventional HPV DNA detection methods. Results The three Raman-active probes showed excellent spectral characteristics with no overlapping characteristic peaks. The SVM model achieved an overall classification accuracy of 93.0%, with genotyping accuracies of 95.2%, 93.7%, and 91.4% for HPV16, HPV18, and other high-risk HPV types, respective-ly. ROC curve analysis showed AUC values of 0.978, 0.962, and 0.943. The Raman spectroscopy method exhibited a sensitivity of 92.5%, specificity of 94.3%, accuracy of 93.3%, with a Kappa coefficient of 0.865. The detection limit for high-risk HPV was 0.1 IU/mL with good reproducibility (coefficient of variation: 4.3%). No significant difference was observed in temperature sta-bility tests (4~37 ℃, P>0.05). Interference tests indicated signal deviations for hemoglobin and albumin were within 8% and 6%, respectively, while those for IL-6 and TNF-α were below 7% and 6%. Continuous 30-day monitoring indicated the Raman signal intensity decay rate was 3.8%, indicating good stability. The detection time was shortened to 45.3±6.7 minutes, and cost decreased to 4 yuan per sample. Conclusions The high-risk HPV detection method based on Raman spectroscopy has good sen-sitivity, specificity, and genotyping capability. Compared with traditional methods, it offers has advantages such as shorter detec-tion time, lower cost, and higher specificity, presenting broad clinical application prospects.

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备注/Memo

备注/Memo:
基金项目：江苏省宿迁市科技指令性项目( S202312 )。
作者简介：葛敏 (1986-)，女，学士，副主任技师，主要从事拉曼光谱及免疫学相关研究，E-mail：donwenjun@126.com。
通讯作者：裴兵 (1972-)，男，硕士研究生，主任技师，主要从事分子免疫及实验室管理研究，E-mail：sqpeibing@njmu.edu.cn。
更新日期/Last Update: 2026-03-15