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Genotyping of 30 kinds of cutaneous human papillomaviruses by a multiplex microfluidic loop-mediated isothermal amplification and visual detection method.
Virol J. 2020 07 09; 17(1):99.VJ

Abstract

BACKGROUND

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.

OBJECTIVES

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.

METHODS

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.

RESULTS

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.

CONCLUSIONS

The new cutaneous type HPV detection system is characterized by both a good sensitivity and specificity compared to conventional methods.

Authors+Show Affiliations

Department of Dermatology, The First Hospital of China Medical University, No.155 Nanjing Bei Street, Heping District, Shenyang, Liaoning Province, 110001, PR China. NHC Key Laboratory of Immunodermatology (China Medical University), No.155 Nanjing Bei Street, Heping District, Shenyang, Liaoning Province, 110001, PR China. Key Laboratory of Immunodermatology (China Medical University), Ministry of Education, No.155 Nanjing Bei Street, Heping District, Shenyang, Liaoning Province, 110001, PR China. Key Laboratory of Immunodermatology, Liaoning Province, No.155 Nanjing Bei Street, Heping District, Shenyang, Liaoning Province, 110001, PR China. Key Laboratory of Immunodermatology (China Medical University), Department of education of Liaoning Province, No.155 Nanjing Bei Street, Heping District, Shenyang, Liaoning Province, 110001, PR China.Department of Dermatology, The First Hospital of China Medical University, No.155 Nanjing Bei Street, Heping District, Shenyang, Liaoning Province, 110001, PR China. NHC Key Laboratory of Immunodermatology (China Medical University), No.155 Nanjing Bei Street, Heping District, Shenyang, Liaoning Province, 110001, PR China. Key Laboratory of Immunodermatology (China Medical University), Ministry of Education, No.155 Nanjing Bei Street, Heping District, Shenyang, Liaoning Province, 110001, PR China. Key Laboratory of Immunodermatology, Liaoning Province, No.155 Nanjing Bei Street, Heping District, Shenyang, Liaoning Province, 110001, PR China. Key Laboratory of Immunodermatology (China Medical University), Department of education of Liaoning Province, No.155 Nanjing Bei Street, Heping District, Shenyang, Liaoning Province, 110001, PR China. Key Laboratory of Biopharmaceutical Production & Formulation Engineering, PLA and State Key Laboratory of Biochemical Engineering, Institute of Process Engineering, Chinese Academy of Sciences, Beijing, 100190, PR China.Key Laboratory of Biopharmaceutical Production & Formulation Engineering, PLA and State Key Laboratory of Biochemical Engineering, Institute of Process Engineering, Chinese Academy of Sciences, Beijing, 100190, PR China.Key Laboratory of Biopharmaceutical Production & Formulation Engineering, PLA and State Key Laboratory of Biochemical Engineering, Institute of Process Engineering, Chinese Academy of Sciences, Beijing, 100190, PR China.Dermatology Hospital of Southern Medical University, No. 2 Lujing Road, Yuexiu District, Guangzhou, Guangdong Province, 510091, PR China. sunbarryz@163.com.Key Laboratory of Biopharmaceutical Production & Formulation Engineering, PLA and State Key Laboratory of Biochemical Engineering, Institute of Process Engineering, Chinese Academy of Sciences, Beijing, 100190, PR China. ygdu@ipe.ac.cn.Department of Dermatology, The First Hospital of China Medical University, No.155 Nanjing Bei Street, Heping District, Shenyang, Liaoning Province, 110001, PR China. gaobarry@hotmail.com. NHC Key Laboratory of Immunodermatology (China Medical University), No.155 Nanjing Bei Street, Heping District, Shenyang, Liaoning Province, 110001, PR China. gaobarry@hotmail.com. Key Laboratory of Immunodermatology (China Medical University), Ministry of Education, No.155 Nanjing Bei Street, Heping District, Shenyang, Liaoning Province, 110001, PR China. gaobarry@hotmail.com. Key Laboratory of Immunodermatology, Liaoning Province, No.155 Nanjing Bei Street, Heping District, Shenyang, Liaoning Province, 110001, PR China. gaobarry@hotmail.com. Key Laboratory of Immunodermatology (China Medical University), Department of education of Liaoning Province, No.155 Nanjing Bei Street, Heping District, Shenyang, Liaoning Province, 110001, PR China. gaobarry@hotmail.com.Department of Dermatology, The First Hospital of China Medical University, No.155 Nanjing Bei Street, Heping District, Shenyang, Liaoning Province, 110001, PR China. xiaoqiliumin@163.com. NHC Key Laboratory of Immunodermatology (China Medical University), No.155 Nanjing Bei Street, Heping District, Shenyang, Liaoning Province, 110001, PR China. xiaoqiliumin@163.com. Key Laboratory of Immunodermatology (China Medical University), Ministry of Education, No.155 Nanjing Bei Street, Heping District, Shenyang, Liaoning Province, 110001, PR China. xiaoqiliumin@163.com. Key Laboratory of Immunodermatology, Liaoning Province, No.155 Nanjing Bei Street, Heping District, Shenyang, Liaoning Province, 110001, PR China. xiaoqiliumin@163.com. Key Laboratory of Immunodermatology (China Medical University), Department of education of Liaoning Province, No.155 Nanjing Bei Street, Heping District, Shenyang, Liaoning Province, 110001, PR China. xiaoqiliumin@163.com.Department of Dermatology, The First Hospital of China Medical University, No.155 Nanjing Bei Street, Heping District, Shenyang, Liaoning Province, 110001, PR China. NHC Key Laboratory of Immunodermatology (China Medical University), No.155 Nanjing Bei Street, Heping District, Shenyang, Liaoning Province, 110001, PR China. Key Laboratory of Immunodermatology (China Medical University), Ministry of Education, No.155 Nanjing Bei Street, Heping District, Shenyang, Liaoning Province, 110001, PR China. Key Laboratory of Immunodermatology, Liaoning Province, No.155 Nanjing Bei Street, Heping District, Shenyang, Liaoning Province, 110001, PR China. Key Laboratory of Immunodermatology (China Medical University), Department of education of Liaoning Province, No.155 Nanjing Bei Street, Heping District, Shenyang, Liaoning Province, 110001, PR China.

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

32646520

Citation

Wang, Yining, et al. "Genotyping of 30 Kinds of Cutaneous Human Papillomaviruses By a Multiplex Microfluidic Loop-mediated Isothermal Amplification and Visual Detection Method." Virology Journal, vol. 17, no. 1, 2020, p. 99.
Wang Y, Ge G, Mao R, et al. Genotyping of 30 kinds of cutaneous human papillomaviruses by a multiplex microfluidic loop-mediated isothermal amplification and visual detection method. Virol J. 2020;17(1):99.
Wang, Y., Ge, G., Mao, R., Wang, Z., Sun, Y. Z., Du, Y. G., Gao, X. H., Qi, R. Q., & Chen, H. D. (2020). Genotyping of 30 kinds of cutaneous human papillomaviruses by a multiplex microfluidic loop-mediated isothermal amplification and visual detection method. Virology Journal, 17(1), 99. https://doi.org/10.1186/s12985-020-01373-3
Wang Y, et al. Genotyping of 30 Kinds of Cutaneous Human Papillomaviruses By a Multiplex Microfluidic Loop-mediated Isothermal Amplification and Visual Detection Method. Virol J. 2020 07 9;17(1):99. PubMed PMID: 32646520.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Genotyping of 30 kinds of cutaneous human papillomaviruses by a multiplex microfluidic loop-mediated isothermal amplification and visual detection method. AU - Wang,Yining, AU - Ge,Ge, AU - Mao,Rui, AU - Wang,Zhuo, AU - Sun,Yu-Zhe, AU - Du,Yu-Guang, AU - Gao,Xing-Hua, AU - Qi,Rui-Qun, AU - Chen,Hong-Duo, Y1 - 2020/07/09/ PY - 2020/02/09/received PY - 2020/06/30/accepted PY - 2020/7/11/entrez PY - 2020/7/11/pubmed PY - 2020/7/11/medline KW - Detection KW - Genotyping KW - HPV KW - LAMP KW - Microfluidic SP - 99 EP - 99 JF - Virology journal JO - Virol. J. VL - 17 IS - 1 N2 - BACKGROUND: 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. OBJECTIVES: 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. METHODS: 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. RESULTS: 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. CONCLUSIONS: The new cutaneous type HPV detection system is characterized by both a good sensitivity and specificity compared to conventional methods. SN - 1743-422X UR - https://www.unboundmedicine.com/medline/citation/32646520/Genotyping_of_30_kinds_of_cutaneous_human_papillomaviruses_by_a_multiplex_microfluidic_loop-mediated_isothermal_amplification_and_visual_detection_method L2 - https://virologyj.biomedcentral.com/articles/10.1186/s12985-020-01373-3 DB - PRIME DP - Unbound Medicine ER -
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