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Development and application of a rapid detection system for human papillomavirus and Herpes simplex virus-2 by loop-mediated isothermal amplification assay.
Microb Pathog. 2016 Aug; 97:178-82.MP

Abstract

Human papillomavirus (HPV) infection is an important factor that causes cervical cancer and non-melanoma skin cancer (NMSC), while HSV-2 plays an important role when HR-HPV triggers the cancer. Thus, a quick and convenient assay in the detection of HPV and HSV-2in the screening of HPV and HSV-2 infection is required. Two respective HPV and HSV-2 detection methods were established based on loop-mediated isothermal amplification (LAMP) assay. Specific outer primers, inner primers, and loop primers were designed according to the conserved domains of HPV and HSV-2 genomes, respectively, while degenerate primers were used for HPV assay. After optimizing the reaction conditions, the results were observed by LAMP Tubidimeter real-time LA-320. Standard plasmids HPV-L-P and HSV-2-L-P were cloned and used in sensitivity tests of HPV LAMP and HSV-2 LAMP, respectively. Fifty samples of actinic keratosis (AK), 20 samples of squamous cell carcinoma (SCC), 50 samples of basal cell carcinoma (BCC) and 20 samples of seborrheic keratosis (SK) were detected by HPV assay. Seventy three clinical samples of vaginitis, chronic cervicitis, cervical intraepithelial neoplasias and cervical cancer level positive were detected with HPV and HSV-2 assays. The reaction conditions of two assays were the same with a reaction temperature of 63 °C and a reaction time of 45 min. The sensitivity of HPV LAMP assay was 10 copies/μL, while that of the HSV-2 LAMP assay was 100 copies/μL. No cross-reactivity was observed. The HPV positive rates of AK, SCC, BCC and SK samples were 80% (40/50), 75% (15/20), 44% (22/50) and 21% (15/72), respectively. As an economic and quick diagnostic tool, LAMP assay is conducive to the extensive screening of HPV and HSV-2 and has huge potential to be promoted in resource-limited hospitals.

Authors+Show Affiliations

Department of Gynaecology and Obstetrics, Binzhou People's Hospital, Binzhou 256610, China.Department of Gynaecology and Obstetrics, Binzhou People's Hospital, Binzhou 256610, China.Department of Gynaecology and Obstetrics, Binzhou Medical University Hospital, Binzhou 256610, China. Electronic address: lukexinedu@sina.com.

Pub Type(s)

Evaluation Study
Journal Article

Language

eng

PubMed ID

27287497

Citation

Yang, Jin-Fang, et al. "Development and Application of a Rapid Detection System for Human Papillomavirus and Herpes Simplex Virus-2 By Loop-mediated Isothermal Amplification Assay." Microbial Pathogenesis, vol. 97, 2016, pp. 178-82.
Yang JF, Zhao CZ, Lu KX. Development and application of a rapid detection system for human papillomavirus and Herpes simplex virus-2 by loop-mediated isothermal amplification assay. Microb Pathog. 2016;97:178-82.
Yang, J. F., Zhao, C. Z., & Lu, K. X. (2016). Development and application of a rapid detection system for human papillomavirus and Herpes simplex virus-2 by loop-mediated isothermal amplification assay. Microbial Pathogenesis, 97, 178-82. https://doi.org/10.1016/j.micpath.2016.06.012
Yang JF, Zhao CZ, Lu KX. Development and Application of a Rapid Detection System for Human Papillomavirus and Herpes Simplex Virus-2 By Loop-mediated Isothermal Amplification Assay. Microb Pathog. 2016;97:178-82. PubMed PMID: 27287497.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Development and application of a rapid detection system for human papillomavirus and Herpes simplex virus-2 by loop-mediated isothermal amplification assay. AU - Yang,Jin-Fang, AU - Zhao,Chang-Zhen, AU - Lu,Ke-Xin, Y1 - 2016/06/08/ PY - 2016/03/06/received PY - 2016/05/31/revised PY - 2016/06/06/accepted PY - 2016/6/12/entrez PY - 2016/6/12/pubmed PY - 2017/3/28/medline KW - Degenerate primers KW - Herpes simplex virus-2 KW - Human papillomavirus KW - Loop-mediated isothermal amplification SP - 178 EP - 82 JF - Microbial pathogenesis JO - Microb. Pathog. VL - 97 N2 - Human papillomavirus (HPV) infection is an important factor that causes cervical cancer and non-melanoma skin cancer (NMSC), while HSV-2 plays an important role when HR-HPV triggers the cancer. Thus, a quick and convenient assay in the detection of HPV and HSV-2in the screening of HPV and HSV-2 infection is required. Two respective HPV and HSV-2 detection methods were established based on loop-mediated isothermal amplification (LAMP) assay. Specific outer primers, inner primers, and loop primers were designed according to the conserved domains of HPV and HSV-2 genomes, respectively, while degenerate primers were used for HPV assay. After optimizing the reaction conditions, the results were observed by LAMP Tubidimeter real-time LA-320. Standard plasmids HPV-L-P and HSV-2-L-P were cloned and used in sensitivity tests of HPV LAMP and HSV-2 LAMP, respectively. Fifty samples of actinic keratosis (AK), 20 samples of squamous cell carcinoma (SCC), 50 samples of basal cell carcinoma (BCC) and 20 samples of seborrheic keratosis (SK) were detected by HPV assay. Seventy three clinical samples of vaginitis, chronic cervicitis, cervical intraepithelial neoplasias and cervical cancer level positive were detected with HPV and HSV-2 assays. The reaction conditions of two assays were the same with a reaction temperature of 63 °C and a reaction time of 45 min. The sensitivity of HPV LAMP assay was 10 copies/μL, while that of the HSV-2 LAMP assay was 100 copies/μL. No cross-reactivity was observed. The HPV positive rates of AK, SCC, BCC and SK samples were 80% (40/50), 75% (15/20), 44% (22/50) and 21% (15/72), respectively. As an economic and quick diagnostic tool, LAMP assay is conducive to the extensive screening of HPV and HSV-2 and has huge potential to be promoted in resource-limited hospitals. SN - 1096-1208 UR - https://www.unboundmedicine.com/medline/citation/27287497/Development_and_application_of_a_rapid_detection_system_for_human_papillomavirus_and_Herpes_simplex_virus_2_by_loop_mediated_isothermal_amplification_assay_ L2 - https://linkinghub.elsevier.com/retrieve/pii/S0882-4010(16)30114-0 DB - PRIME DP - Unbound Medicine ER -