Human 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. To 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. L1 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. The lower detection limit of the LAMP assay was 107 viral 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. The new cutaneous type HPV detection system is characterized by both a good sensitivity and specificity compared to conventional methods.

中文翻译：

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通过多重微流环介导的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。

人乳头瘤病毒（HPV）是一类非包膜的小病毒，带有双链环状DNA，可导致多种皮肤疾病，例如良性疣，在临床上很常见。当前的HPV检测系统主要针对粘膜HPV，但是，缺乏用于皮肤HPV检测的有效临床方法。使用比色环介导的等温扩增（LAMP）和羟基萘酚蓝（HNB）染料结合微流技术，建立皮肤HPV的快速检测系统。化学合成了30种皮肤HPV的L1 DNA序列，并使用在线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，并用肉眼检查扩增结果。使用PCR测序作为对照方法，通过检测临床样本获得了新检测系统的特异性和敏感性。在合成的L1 DNA序列上进行测试时，LAMP分析的下限为107个病毒DNA拷贝/微升，优于常规PCR。与PCR测序相比，HPV27，HPV2，HPV1，HPV57，HPV3，HPV4，HPV7和HPV75基因型检测的敏感性为100％，而在临床样品上检测的特异性分别为34.55、45.12、95.83、98.59和97.62％。 。与传统方法相比，新型皮肤型HPV检测系统的特点是灵敏度和特异性都很高。在合成的L1 DNA序列上进行测试时，LAMP分析的下限为107个病毒DNA拷贝/微升，优于常规PCR。与PCR测序相比，HPV27，HPV2，HPV1，HPV57，HPV3，HPV4，HPV7和HPV75基因型检测的敏感性为100％，而在临床样品上检测的特异性分别为34.55、45.12、95.83、98.59和97.62％。 。与传统方法相比，新型皮肤型HPV检测系统的特点是灵敏度和特异性都很高。在合成的L1 DNA序列上进行测试时，LAMP分析的下限为107个病毒DNA拷贝/微升，优于常规PCR。与PCR测序相比，HPV27，HPV2，HPV1，HPV57，HPV3，HPV4，HPV7和HPV75基因型检测的敏感性为100％，而在临床样品上检测的特异性分别为34.55、45.12、95.83、98.59和97.62％。 。与传统方法相比，新型皮肤型HPV检测系统的特点是灵敏度和特异性都很高。在临床样本上测试时分别为62％。与传统方法相比，新型皮肤型HPV检测系统的特点是灵敏度和特异性都很高。在临床样本上测试时分别为62％。与传统方法相比，新型皮肤型HPV检测系统的特点是灵敏度和特异性都很高。