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| 环介导等温扩增技术对HPV16、58亚型进行分型检测 | |
| 引用本文： | 张可欣1,2,杨思熙1,2,王丽娜1,苗光新3,刘金霞1,邢恩宏4. 环介导等温扩增技术对HPV16、58亚型进行分型检测[J]. 现代预防医学, 2019, 0(20): 3787-3790 |
| 作者姓名： | 张可欣1 2 杨思熙1 2 王丽娜1 苗光新3 刘金霞1 邢恩宏4 |
| 作者单位： | １.承德医学院病原生物学实验中心，河北 承德 067000；2.基础医学研究所，河北 承德 067000；3.承德医学院，河北 承德 067000；4.承德医学院附属医院，河北 承德 067000 |
| 摘 要： | 目的 采用煮沸法处理人宫颈黏液上皮细胞，获得人乳头瘤病毒（HPV）DNA，然后再通过应用可视化环介导等温扩增（LAMP）检测技术对 HPV常见基因型16、58的2个亚型进行快速诊断，探讨 HPV感染和分型的快速检测技术，以指导 HPV感染患者的临床诊断和治疗。方法 收集承德医学院妇科宫颈脱落细胞标本104例，采用直接煮沸法提取HPV DNA。应用LAMP及聚合酶链式反应（PCR）2种诊断技术进行检测，并在LAMP扩增体系中加入钙黄绿素使产物可视化。对检测结果进行特异性和敏感度比较，并计算 kappa系数对校测结果的一致性进行分析。结果 成功建立了 HPV16、58的可视化 LAMP体系，其特异性较好，且钙黄绿素产物与电泳结果一致， HPV16、58 LAMP检测限分别为100拷贝/管和103拷贝/管，与 PCR一致性较好（kappa值＞0.7）且无统计学差异（P＞0.05）。结论 LAMP对未经纯化的 HPV核酸具有较好特异性和敏感度，且操作简单，适合应用于基层。 |
| 关 键 词： | 人乳头瘤病毒（HPV） 可视化环介导等温扩增（LAMP） 分型检测 快速检测 |
Loop-mediated isothermal amplification for detecting and typing of human papillomavirus type 16 and 58 |
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| ZHANG Ke-xin,YANG Si-xi,WANG Li-na,MIAO Guang-xin,LIU Jin-xia,XING En-hong. Loop-mediated isothermal amplification for detecting and typing of human papillomavirus type 16 and 58[J]. Modern Preventive Medicine, 2019, 0(20): 3787-3790 | |
| Authors: | ZHANG Ke-xin YANG Si-xi WANG Li-na MIAO Guang-xin LIU Jin-xia XING En-hong |
| Affiliation: | *Chengde Medical University, Chengde Hebei 067000, China |
| Abstract: | Objective The aim of this study was to perform rapid diagnosis of HPV16 and HPV 58 by visual loop-mediated isothermal amplification (LAMP) using human papillomavirus (HPV) DNA obtained by boiling human cervical mucoepithelial cells, in order to explore the rapid detection technology of HPV infection and typing, as to guide the clinical diagnosis and treatment of HPV infection patients. Methods A total of 104 cases of cervical ablated cells were collected, and HPV DNA was extracted by direct boiling method. Two diagnostic techniques, LAMP and polymerase chain reaction(PCR), were used for detection. The product was visualized by adding calcein in LAMP amplification system. The specificity and sensitivity of test results were compared. Kappa coefficient was calculated to analyze the consistency of test results. Results The visual LAMP system of HPV16 and 18 was successfully established. The specificity was good. The product of calcitrochlorin was consistent with the electrophoresis results. The detection limits of LAMP of HPV16 and 58 were 100 copies/tube and 103 copies/tube, respectively, which were consistent with PCR (Kappa value of >0.7). There was no significant difference (P>0.05). Conclusion The LAMP had good specificity and sensitivity towards unpurified HPV nucleic acid, and was easy to manipulate. LAMP can be used in the basic medical units. |
| Keywords: | Human papillomavirus (HPV) Visual loop-mediated isothermal amplification (LAMP) Genotyping detection Rapid detection |
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