期刊论文详细信息
Brazilian Journal of Infectious Diseases
Human papillomavirus detection and typing using a nested-PCR-RFLP assay
Janaina Coser2  Thaís Da Rocha Boeira1  André Salvador Kazantzi Fonseca1  Nilo Ikuta2  Vagner Ricardo Lunge2 
[1] ,Universidade Luterana do Brasil Postgraduate Program in Genetics and Molecular Diagnosis Canoas RS ,Brazil
关键词: DNA probes;    HPV;    polymerase chain reaction;    polymorphism;    restriction;    fragment length;   
DOI  :  10.1590/S1413-86702011000500009
来源: SciELO
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【 摘 要 】

BACKGROUND: It is clinically important to detect and type human papillomavirus (HPV) in a sensitive and specific manner. OBJECTIVES: Development of a nested-polymerase chain reaction-restriction fragment length polymorphism (nested-PCR-RFLP) assay to detect and type HPV based on the analysis of L1 gene. METHODS: Analysis of published DNA sequence of mucosal HPV types to select sequences of new primers. Design of an original nested-PCR assay using the new primers pair selected and classical MY09/11 primers. HPV detection and typing in cervical samples using the nested-PCR-RFLP assay. RESULTS: The nested-PCR-RFLP assay detected and typed HPV in cervical samples. Of the total of 128 clinical samples submitted to simple PCR and nested-PCR for detection of HPV, 37 (28.9%) were positive for the virus by both methods and 25 samples were positive only by nested-PCR (67.5% increase in detection rate compared with single PCR). All HPV positive samples were effectively typed by RFLP assay. CONCLUSION: The method of nested-PCR proved to be an effective diagnostic tool for HPV detection and typing.

【 授权许可】

CC BY-NC-ND   
 All the contents of this journal, except where otherwise noted, is licensed under a Creative Commons Attribution License

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