Background: Human papillomaviruses (HPV) are implicated in cervical cancer and, recently in oral cancer. In Mexico, there are few studies on oral cancer, therefore the interest in identifying the HPV frequency of low ...Background: Human papillomaviruses (HPV) are implicated in cervical cancer and, recently in oral cancer. In Mexico, there are few studies on oral cancer, therefore the interest in identifying the HPV frequency of low and high risk in samples of the oral and cervical cavities, and determining some risk factors. Objective: To determine the frequency of high and low risk HPV infection in the oral cavity of women with cervical HPV, and to correlate the infection site with risk factors. Materials and Methods: Eighteen female patients between 24 and 53 years, with antecedents of genital HPV infection were included. Both samples of oral cavity and cervix were obtained. DNA extraction from the epithelial cells was performed using the Qiagen kit. PCR was done and the amplicon was observed in 2% agarose gels stained with ethidium bromide. A correlation of HPV infection and risk factors was done. Results: HPV-DNA was detected in the 67% of both samples. The frequency of oral and cervix low risk HPV-DNA was 50%, while high risk HPV-DNA in oral cavity was detected in 17%, and 39% in the cervix. The study of the risk factors involved in HPV infection showed that the participants had the habits of smoking 39%;alcohol drinking 28%;and 78% oral sex. Conclusion: The results showed a high frequency of HPV (67%) infection in the oral and genital mucosas, suggesting that patient’s habits could contribute to the infection;the presence of HPV in the oral mucosa may act as reservoir for new HPV infections.展开更多
Here, we report a novel and universal methodology,termed ＂ntarctic thermolabile uracil-DNA-glycosylase （AUDG）-supplemented nucleic acid amplification techniques （NAAs） using a labeled-based nanoparticle lateral f...Here, we report a novel and universal methodology,termed ＂ntarctic thermolabile uracil-DNA-glycosylase （AUDG）-supplemented nucleic acid amplification techniques （NAAs） using a labeled-based nanoparticle lateral flow biosensor （LFB）＂ （AUDG-NAAs-LFB）, which merges enzymatic （AUDG） digestion of contaminant amplicons with different nucleic acid amplification techniques （NAAs）, and uses a lateral flow biosensor （LFB） for the rapid and visual confirmation of the presence of a target nucleic acid sequence. AUDG-NNAs-LFB is a one-pot, closedvessel assay, that can effectively eliminate false-positive signals arising from either carryover contaminants or the interaction between labeled primers. A new LFB was devised for detecting three targets （two amplicons generated from amplification of target sequences, and a chromatography control）, without the need for probe- hybridization or additional incubation steps. As a proof of concept, multiple cross displacement amplification （MCDA）, which is a specific, sensitive, and rapid isothermal amplification method, was selected as the model amplification technique to demonstrate the feasibility of AUDG-NAAs-LFB. As a result, we demonstrate the applicability of the AUDG-MCDA-LFB method for simultaneously detecting high-risk human papillomaviruses genotypes 16 and 18, which are the most and second-most prevalent strains of the virus reported in women worldwide. We also confirm the principle behind the AUDG-MCDA- LFB assay and validate its sensitivity, reproducibility, and specificity using serial dilutions of the type-specific plasmids, as well as clinical samples. This proof- of-concept method （AUDG-MCDA-LFB） can be easily reconfigured to detect various nudeic acid sequences by redesigning the specific MCDA primers.展开更多
文摘Background: Human papillomaviruses (HPV) are implicated in cervical cancer and, recently in oral cancer. In Mexico, there are few studies on oral cancer, therefore the interest in identifying the HPV frequency of low and high risk in samples of the oral and cervical cavities, and determining some risk factors. Objective: To determine the frequency of high and low risk HPV infection in the oral cavity of women with cervical HPV, and to correlate the infection site with risk factors. Materials and Methods: Eighteen female patients between 24 and 53 years, with antecedents of genital HPV infection were included. Both samples of oral cavity and cervix were obtained. DNA extraction from the epithelial cells was performed using the Qiagen kit. PCR was done and the amplicon was observed in 2% agarose gels stained with ethidium bromide. A correlation of HPV infection and risk factors was done. Results: HPV-DNA was detected in the 67% of both samples. The frequency of oral and cervix low risk HPV-DNA was 50%, while high risk HPV-DNA in oral cavity was detected in 17%, and 39% in the cervix. The study of the risk factors involved in HPV infection showed that the participants had the habits of smoking 39%;alcohol drinking 28%;and 78% oral sex. Conclusion: The results showed a high frequency of HPV (67%) infection in the oral and genital mucosas, suggesting that patient’s habits could contribute to the infection;the presence of HPV in the oral mucosa may act as reservoir for new HPV infections.
文摘Here, we report a novel and universal methodology,termed ＂ntarctic thermolabile uracil-DNA-glycosylase （AUDG）-supplemented nucleic acid amplification techniques （NAAs） using a labeled-based nanoparticle lateral flow biosensor （LFB）＂ （AUDG-NAAs-LFB）, which merges enzymatic （AUDG） digestion of contaminant amplicons with different nucleic acid amplification techniques （NAAs）, and uses a lateral flow biosensor （LFB） for the rapid and visual confirmation of the presence of a target nucleic acid sequence. AUDG-NNAs-LFB is a one-pot, closedvessel assay, that can effectively eliminate false-positive signals arising from either carryover contaminants or the interaction between labeled primers. A new LFB was devised for detecting three targets （two amplicons generated from amplification of target sequences, and a chromatography control）, without the need for probe- hybridization or additional incubation steps. As a proof of concept, multiple cross displacement amplification （MCDA）, which is a specific, sensitive, and rapid isothermal amplification method, was selected as the model amplification technique to demonstrate the feasibility of AUDG-NAAs-LFB. As a result, we demonstrate the applicability of the AUDG-MCDA-LFB method for simultaneously detecting high-risk human papillomaviruses genotypes 16 and 18, which are the most and second-most prevalent strains of the virus reported in women worldwide. We also confirm the principle behind the AUDG-MCDA- LFB assay and validate its sensitivity, reproducibility, and specificity using serial dilutions of the type-specific plasmids, as well as clinical samples. This proof- of-concept method （AUDG-MCDA-LFB） can be easily reconfigured to detect various nudeic acid sequences by redesigning the specific MCDA primers.
