Development and validation of a method for human papillomavirus genotyping based on molecular beacon probes

Autor: Yu Yuefeng, Ren Xuyi, Pan Caixia, Jiangfeng Lyu, Jing Zhou
Rok vydání: 2018
Předmět:
0301 basic medicine
Viral Diseases
Genotyping Techniques
Artificial Gene Amplification and Extension
Pathology and Laboratory Medicine
Cervical Cancer
Polymerase Chain Reaction
law.invention
Human Papillomavirus DNA Tests
law
Genotype
Medicine and Health Sciences
DNA Probes
HPV
Papillomaviridae
Polymerase chain reaction
Cervical cancer
Multidisciplinary
Hybridization probe
Physics
Melting
Condensed Matter Physics
Infectious Diseases
Molecular Diagnostic Techniques
Oncology
Medical Microbiology
Viral Pathogens
Viruses
Physical Sciences
Medicine
Pathogens
Phase Transitions
Research Article
Human Papillomavirus Infection
Genotyping
Papillomaviruses
HPV-6
Science
Urology
030106 microbiology
Sexually Transmitted Diseases
Molecular Probe Techniques
Biology
Research and Analysis Methods
Sensitivity and Specificity
Microbiology
Melting curve analysis
03 medical and health sciences
Molecular beacon
medicine
Humans
Human papillomavirus
Molecular Biology Techniques
Microbial Pathogens
Molecular Biology
Biology and life sciences
Genitourinary Infections
Papillomavirus Infections
Organisms
Human Papillomavirus
Cancers and Neoplasms
medicine.disease
Virology
Probe Hybridization
DNA viruses
Gynecological Tumors
Zdroj: PLoS ONE
PLoS ONE, Vol 13, Iss 11, p e0207930 (2018)
ISSN: 1932-6203
Popis: We describe a new assaying system for the detection and genotyping of human papillomavirus (HPV) based on linear-after-the-exponential-PCR(LATE-PCR) and melting curve analysis. The 23 most prevalent HPV strains (types 6, 11, 16, 18, 31, 33, 35, 39, 42, 45, 51, 52, 53, 56, 58, 59, 66, 68, 70, 73, 81, 82, and 83) are assayed in two sealed reaction tubes within 2 h. Good sensitivity and specificity was evaluated by testing cloned HPV DNA and clinical samples. The detection limit was 5-500 copies/reaction depending on the genotype. No cross-reactivity was observed with the other HPV types that are not covered by our method or pathogens tested which were commonly found in female genital tract. When compared with the HPV GenoArray Diagnostic kit, the results from 1104 clinical samples suggest good overall agreement between the two methods,(98.37%, 95% CI: 97.44%-98.97%) and the kappa value was 0.954. Overall, this new HPV genotyping assay system presents a simple, rapid, universally applicable, sensitive, and highly specific detection methodology that should be useful for HPV detection and genotyping, therefore, is potentially of great value in clinical application.
Databáze: OpenAIRE
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