Human papillomavirus detection and typing using a nested-PCR-RFLP assay

Coser, Janaina; Boeira, Thaís da Rocha; Fonseca, André Salvador Kazantzi; Ikuta, Nilo; Lunge, Vagner Ricardo

doi:10.1016/s1413-8670(11)70229-x

Cited by 18 publications

(18 citation statements)

References 28 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…The high prevalence of oncogenic genotypes found in this study indicates the need for frequent monitoring, and suggests nPCR technique for DNA-HPV detection as an alternative to early identification of women at high risk for cervical cancer development. Previous studies have already demonstrated that the use of primers pair MY09/11 and GP5/GP6 in a nPCR assay increases the sensitivity of HPV detection compared with PCR assay (Seaman et al , 2010; Coser et al , 2011). …”

Section: Discussionmentioning

confidence: 99%

High incidence of oncogenic HPV genotypes found in women from Southern Brazil

Entiauspe¹,

Silveira²,

Nunes³

et al. 2014

Braz. J. Microbiol.

View full text Add to dashboard Cite

Oncogenic HPV genotypes are strongly associated with premalignant and malignant cervical lesion. The purpose was to determine human papillomavirus (HPV) prevalence and genotypes, and to estimate cervical cancer risk factor associations. Cervical samples were obtained from 251 women seeking gynecological care at the Pelotas School of Medicine Clinic. This is a cross-sectional study. HPV-DNA was amplified by nested-PCR using MY09/11 and GP5/6 primers, and the sequencing was used for genotyping. Sociodemographic and behavioral risk factors were obtained by closed questionnaire, and its relationship to HPV infection prevalence were analyzed. Statistical analyses were performed using SPSS 16.0 software, and differences were considered significant at p < 0.05. As results, the prevalence of HPV infection was 29.9%. The most frequent genotype was HPV-16 (41.3%), followed by HPV-18 (17.3%), and HPV-33 (9.3%). Others nine HPV genotypes were also found. On this population, prevalence of oncogenic HPV genotypes was high, but does not seem to confer relationship with the risk factors investigated. Future investigations in larger populations are necessary, for the proposition of more appropriated monitoring strategies and treatment according to the Brazilian health service reality, as well as patients.

show abstract

Section: Discussionmentioning

confidence: 99%

High incidence of oncogenic HPV genotypes found in women from Southern Brazil

Entiauspe¹,

Silveira²,

Nunes³

et al. 2014

Braz. J. Microbiol.

View full text Add to dashboard Cite

show abstract

“…HPV DNA samples were analyzed by first applying SB01/02 PCR primers ( Table 1 ) according to Coser et al [ 29 ] To increase the HPV detection rate the assay was modified and optimized for template concentration (3 μl in a final volume of 20 μl reaction), annealing temperature (42 °C), duration of each step (as indicated in Table 2 ), number of cycles (from 20 to 40 in two steps), and the MgCl 2 concentration that was adjusted to 3.5 mM. Thermal cycling conditions for outer PCR ( Table 2 ) were modified from Coser et al [ 29 ]. PCR product was approximately 495 bp for the majority of HPV genotypes.…”

Section: Methodsmentioning

confidence: 99%

Molecular detection of human papillomavirus (HPV) in highly fragmented DNA from cervical cancer biopsies using double-nested PCR

et al. 2018

View full text Add to dashboard Cite

show abstract

“…The primers target conserved regions of the HPV genome, such as the L1 capsid gene [37]. After amplification, the HPV genotypes can be determined separately, using techniques such as restriction-fragment length polymorphism (RFLP), linear probe assays, direct sequencing, or genotype-specific primers [38]. Some researchers have used a type-specific PCR, with primers that amplify the long control region L1 and E6/E7 [39].…”

Section: Introductionmentioning

confidence: 99%

“…Amplification of samples containing DNA from more than one HPV genotype can lead to a much stronger amplification of one of the sequences present, which would complicate the detection of all genotypes in a sample with multiple infections. Sometimes, additional, labor-intensive procedures, such as sequencing or type-specific PCR, are required [38]. …”

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

A review of methods for detect human Papillomavirusinfection

Abreu

Souza

Gimenes

et al. 2012

Virol J

230

197

View full text Add to dashboard Cite

Human Papillomavirus (HPV) is the most common sexually transmitted virus. Worldwide, the most common high-risk (HR)-HPV are -16/18, and approximately 70% of cervical cancers (CC) are due to infection by these genotypes. Persistent infection by HR-HPV is a necessary but not sufficient cause of this cancer, which develops over a long period through precursor lesions, which can be detected by cytological screening. Although this screening has decreased the incidence of CC, HPV-related cervical disease, including premalignant and malignant lesions, continues to be a major burden on health-care systems. Although not completely elucidated, the HPV-driven molecular mechanisms underlying the development of cervical lesions have provided a number of potential biomarkers for both diagnostic and prognostic use in the clinical management of women with HPV-related cervical disease, and these biomarkers can also be used to increase the positive predictive value of current screening methods. In addition, they can provide insights into the biology of HPV-induced cancer and thus lead to the development of nonsurgical therapies. Considering the importance of detecting HPV and related biomarkers, a variety of methods are being developed for these purposes. This review summarizes current knowledge of detection methods for HPV, and related biomarkers that can be used to discriminate lesions with a high risk of progression to CC.

show abstract

Human papillomavirus detection and typing using a nested-PCR-RFLP assay

Abstract: The method of nested-PCR proved to be an effective diagnostic tool for HPV detection and typing.

Cited by 18 publications

References 28 publications

High incidence of oncogenic HPV genotypes found in women from Southern Brazil

High incidence of oncogenic HPV genotypes found in women from Southern Brazil

Molecular detection of human papillomavirus (HPV) in highly fragmented DNA from cervical cancer biopsies using double-nested PCR

A review of methods for detect human Papillomavirusinfection

Contact Info

Product

Resources

About