Tipagem e estado físico de papilomavírus humano por hibridização in situ em lesões intra-epiteliais do colo uterino

Bagarelli, Lúcia Buchalla; Oliani, António Hélio

doi:10.1590/s0100-72032004000100009

Cited by 5 publications

(3 citation statements)

References 15 publications

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“…Molecular diagnosis of HPV infection is important for virus screening, and is mainly based on methods such as: hybrid capture (CH2) (20,23) , in situ hybridization (2) , and PCR (8,21,24) . These techniques have vary widely in terms of sensitivity and specificity, and PCR is the most used today in various areas of molecular diagnostics due to its great ability to detect small fragments of deoxyribonucleic acid (DNA) (4,11) .…”

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confidence: 99%

HPV detection using primers MY09/MY11 and GP5+/GP6+ in patients with cytologic and/or colposcopic changes

Venceslau¹,

Bezerra²,

Lopes³

et al. 2014

Jornal Brasileiro De Patologia E Medicina Laboratorial

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Introduction: Cervical cancer is one of the most common diseases among women, and cause considerable morbidity and mortality. Considering that cervical cancer is an important neoplasia in northeastern Brazil, and the prevalence of high-risk human papillomavirus (HPV) is directly associated with it, this work had aimed to correlate the cytological and/or colposcopic findings with HPV infection status, and verify the performance of MY09/MY11 and GP5+/6+ primers for HPV detection. Material and method: Patients in this study were from Penedo-AL, a city with high level of poverty (poverty rate of 60.62%). Out of 70 patients with cytological and/or colposcopic changes, 32 agreed to participate in the study. Results: Regarding cytology, 21 (30%) patients presented atypical squamous cells of undetermined significance (ASC-US); 20 (29%), atypical glandular cells of undetermined significance (AGUS); 12 (17%), low-grade intraepithelial lesion (LSIL); five (7%), high-grade intraepithelial lesion (HSIL); and 12 (17%), positive colposcopy. From these, 27 (84%) presented the band gene encoding for human ß-globin. From the 27 patients, eight (30%) were positive for HPV. The results showed that the deoxyribonucleic acid (DNA) of HPV was detected in 15% and 30% by using MY-PCR and GP +-PCR, respectively. Conclusion: This study suggests that more than one type of oligonucleotide primer should be used in clinical samples to increase sensitivity for the detection of HPV.

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confidence: 99%

HPV detection using primers MY09/MY11 and GP5+/GP6+ in patients with cytologic and/or colposcopic changes

Venceslau¹,

Bezerra²,

Lopes³

et al. 2014

Jornal Brasileiro De Patologia E Medicina Laboratorial

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“…With 4,800 fatalities and 18,430 new cases , cervical cancer is the fourth leading cause of death in women in Brazil. The molecular diagnostic techniques such as hybrid capture, in situ hybridization, among others, have been a major advances in early diagnosis of cervical cancer associated to HPV. Currently 44% of cases are cancer precursor lesion, called in situ or noninvasive, can be completely cured if treated with such techniques.…”

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confidence: 99%

A Simple HPV 18 Detection Method Based on Ultra Specific Primer Immobilized on Glass Slides

Fidelis

Dutra

Souza

et al. 2013

Clinical Laboratory Analysis

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This study was carried out to develop a simple and inexpensive method for detection of Human papillomavirus (HPV 18) based on irreversible immobilization of ultra specific primer on silanized glass slides. This method is revealed by Blue Green Loading Dye I (LGC) and compared with conventional polymerase chain reaction (PCR) for endocervical samples. The new method was tested in 40 DNA samples with precancer uterine lesions of women treated in Hospital of Recife PE, Brazil. DNA samples were extracted using Wizard Genomic DNA Purification Kit according to manufacturer's instructions. The samples were tested for HPV 18 by conventional PCR (PCRc) and the products visualized on 1.2% agarose gel, with LGC under UV 260 nm. After that the positive and negative samples to HPV 18 were tested by immobilization method and the results visualized with LGC under UV 260 nm. Both PCRc and immobilization method showed high degree of correlation (95%), whereas comparison between PCRc and immobilization method showed good correlation (100%). PCRc is widely known for detection of HPV because of its high sensitivity and efficiency, but due to high cost it is not yet standardized for use in public health laboratories. In our study, the single-stranded DNA immobilized method on a glass slide was effective in screening for HPV revealed by Blue Green and may be an alternative method for diagnosis of HPV once it offers a fast, and easy handling.

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“…Entre eles, a captura híbrida 2 (CH2) é o método molecular mais utilizado em nosso meio para a detecção de HPV. Esta técnica baseia-se na hibridização de DNA, fazendo uso de sondas específicas contra os tipos de HPV considerados de alto risco (1,15) . A PCR baseia-se na amplificação específica de segmentos do DNA alvo e tem potencial para a detecção de níveis muito baixos de carga viral em células e tecidos, mesmo em infecções ditas não produtivas (26) .…”

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Comparação das técnicas de captura de híbridos e PCR para a detecção de HPV em amostras clínicas

Rodrigues¹,

Cantarelli²,

Frantz³

et al. 2009

J. Bras. Patol. Med. Lab.

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Introdução e objetivos: São conhecidos mais de 100 tipos de papilomavírus humano (HPV), dos quais 30 têm sido reportados em infecções anogenitais. A infecção tem importância clínica, pois alguns tipos virais estão associados a lesões que podem progredir para o câncer cervical. Sabe-se que os métodos moleculares são muito importantes para o diagnóstico dessa infecção. O objetivo do estudo é comparar a detecção de HPV de alto risco pelo método de captura híbrida 2 (CH2) com a detecção do vírus pela reação em cadeia da polimerase convencional (PCRc) e em tempo real (PCR-TR). Metodologia: Foram analisadas 56 amostras ectocervicais por CH2 e, após, por PCRc e PCR-TR. Resultados: Ambas, PCRc e PCR-TR, apresentaram alta concordância entre si (95,1%), enquanto a comparação entre as PCRs e a CH2 mostrou concordância razoável entre os resultados (PCRc = 90,2% e PCR-TR = 87,8%). Discussão e conclusão: A CH é aceita para a detecção do HPV, entretanto pode ser menos sensível em comparação com as técnicas de PCR. A PCR-TR tem a vantagem sobre a PCRc em termos de velocidade, sendo também um pouco mais sensível. Devido à alta sensibilidade e à rapidez, os métodos de PCR poderiam ser usados para a triagem de HPV em amostras ectocervicais.

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Tipagem e estado físico de papilomavírus humano por hibridização in situ em lesões intra-epiteliais do colo uterino

Cited by 5 publications

References 15 publications

HPV detection using primers MY09/MY11 and GP5+/GP6+ in patients with cytologic and/or colposcopic changes

HPV detection using primers MY09/MY11 and GP5+/GP6+ in patients with cytologic and/or colposcopic changes

A Simple HPV 18 Detection Method Based on Ultra Specific Primer Immobilized on Glass Slides

Comparação das técnicas de captura de híbridos e PCR para a detecção de HPV em amostras clínicas

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