Abstract
Introduction: a sensitive method of detection for Human Papillomavirus (HPV) is important to facilitate the early treatment of cervical cancer precursors. Objective: to analyze the spectrum of HPV infection and compare the sensibility of DNA HPV detection using polymerase chain reaction (PCR) and nested PCR (nPCR) methods in a group of 251 women of Pelotas-RS. Methods: genomic DNA was extracted from the collected samples and was submitted to PCR methods with the primers MY09/11 and nPCR with the pair of primers MY09/MY11 and GP5+/6+. The results were applied to the softwares EpiInfo v.3.5.1® and STATA v.11 ® for analyzes. Results: the prevalence of HPV infection was 6.8% with the use of primers MY09/11. When associated with primers GP5/6, this result increased to 29.9% (p < 0.001). Conclusion: the increase founded in HPV DNA detection from 6.8 to 29.9% suggests that the technique of nPCR MY09/11 followed by GP5/6 is the most sensitive method to detect HPV DNA from cervical specimens.
Highlights
Introduction: a sensitive method of detection for Human Papillomavirus (HPV) is important to facilitate the early treatment of cervical cancer precursors
All 251 samples were amplified for the TP53 gene, checking DNA quality for HPV detection by polymerase chain reaction (PCR) and nested PCR (nPCR) methods
The HPV-DNA analysis by PCR MY09/11 was observed in 17 samples, whereas nPCR technique found HPV positivity in more 58 samples (24.8%) shown in Table 2, increasing up to 4 times the detection of viral DNA (p < 0.001)
Summary
Introduction: a sensitive method of detection for Human Papillomavirus (HPV) is important to facilitate the early treatment of cervical cancer precursors. With the introduction of biomolecular techniques, it was possible to confirm that the cervical cancer (CC) development is closely associated with the HPV, showing the significance of the HPV infection for the dysplasia development and the transformation of normal cervical cells into cancerous[3,4] This type of cancer is the responsible for the death of 31.400 women in Latin America, 11.000 only in Brazil[5,6] each year. A variation of this technique called nested-PCR (nPCR) with the MY09/11 and GP5+/6+ primer sets, is a high sensitive specific method for HPV DNA detection[11], whith both targeting the L1 conserved region of the viral genome, allowing the detection of a broad range of HPV types[12]. The GP5 and GP6 primer set consists of a fixed nucleotide sequence for each primer and detects a wide range of HPV types by using a lowered annealing temperature during PCR, and targets a 140bp sequence of HPV L1 gene, located inside the sequence recognized by the MY primers[12]
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