Genotyping of 30 kinds of cutaneous human papillomaviruses by a multiplex microfluidic loop-mediated isothermal amplification and visual detection method
Genotyping of 30 kinds of cutaneous human papillomaviruses by a multiplex microfluidic loop-mediated isothermal amplification and visual detection method
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多重微流控环介导等温扩增和视觉检测方法对30种皮肤人乳头瘤病毒进行基因分型
DOI：
10.1186/s12985-020-01373-3
复制
发表时间：
2020-07
期刊：
影响因子：
--
通讯作者：
Chen HD
中科院分区：
文献类型：
--
作者：
Wang Y;Ge G;Mao R;Wang Z;Sun YZ;Du YG;Gao XH;Rui-Qun Qi;Chen HD
研究背景人乳头瘤病毒（Human papillomavirus，HPV）是一组无包膜的双链环状DNA小病毒，可引起良性疣等多种皮肤病，临床上十分常见。目的建立一种以羟基萘酚蓝（HNB）为染料的环介导等温扩增（LAMP）技术结合微流控技术快速检测皮肤HPV的方法。利用在线LAMP设计工具设计了针对L1 DNA的LAMP引物。使用水浴进行等温扩增，用肉眼检查扩增结果。结果LAMP方法对合成的L1 DNA序列的检测下限为107拷贝/μl，优于常规PCR方法。与PCR测序相比，HPV 27、HPV 2、HPV 1、HPV 57、HPV 3、HPV 4、HPV 7和HPV 75基因型检测的敏感性均为100%，特异性分别为34.55、45.12、95.83、98.59和97.62%。结论新的皮肤型HPV检测系统与传统方法相比具有良好的敏感性和特异性。
BackgroundHuman papillomaviruses (HPVs), a group of non-enveloped small viruses with double-stranded circular DNA which lead to multiple skin diseases such as benign warts, are commonly seen in clinics. The current HPV detection systems aim mainly at mucosal HPVs, however, an efficient clinical approach for cutaneous HPVs detection is lacking.ObjectivesTo establish a rapid detection system for cutaneous HPVs using a colorimetric loop-mediated isothermal amplification (LAMP) with hydroxynaphthol blue (HNB) dye in combination with microfluidic technology.MethodsL1 DNA sequences of the 30 cutaneous HPVs were chemically synthesized, and LAMP primers against L1 DNA were designed with use of an online LAMP designing tool. Isothermal amplification was performed with use of a water bath and the amplification results were inspected with the naked eye. Using PCR sequencing as a control method, the specificity and sensitivity of the new detection system were obtained by detecting clinical samples.ResultsThe lower detection limit of the LAMP assay was 107viral DNA copies/μl when tested on synthesized L1 DNA sequences, which was better than the conventional PCR. Compared to PCR sequencing, the sensitivity of HPV27, HPV2, HPV1, HPV57, HPV3, HPV4, HPV7 and HPV75 genotypes detections were 100%, whereas the specificity was 34.55, 45.12, 95.83, 98.59 and 97.62% respectively, when tested on clinical samples.ConclusionsThe new cutaneous type HPV detection system is characterized by both a good sensitivity and specificity compared to conventional methods.
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影响因子：
16.6
作者：
Zheng, B;Ismagilov, RF
通讯作者：
Ismagilov, RF
DOI：
--
发表时间：
2016-09
期刊：
--
影响因子：
--
作者：
Hu Zongyue;Xu Zhouheng;Lu Yiyu
通讯作者：
Hu Zongyue;Xu Zhouheng;Lu Yiyu
影响因子：
5.3
作者：
通讯作者：
--
影响因子：
0.7
作者：
Junxiao Lin;B. Ma;Jiehong Fang;Ye-Fu Wang;Haizhen He;Wei-Yee Lin;Wei Su;Mingzhou Zhang
通讯作者：
Junxiao Lin;B. Ma;Jiehong Fang;Ye-Fu Wang;Haizhen He;Wei-Yee Lin;Wei Su;Mingzhou Zhang
影响因子：
4
作者：
Rocky E Bacelieri;Sandra M. Johnson
通讯作者：
Rocky E Bacelieri;Sandra M. Johnson