Expanded HPV Genotyping by Single-Tube Nested-Multiplex PCR May Explain HPV-Related Disease Recurrence
The role of the human papillomavirus (HPV) in the establishment of cervical cancer has driven studies to find more effective methods of viral detection so that early intervention strategies can be performed. However, the methods still have limitations, especially regarding detecting the different ge...
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2024-11-01
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| author | Luiz Ricardo Goulart Bruna França Matias Colombo Mayara Ingrid Sousa Lima Maria Socorro A. de Andrade Juliana São Julião Adriana Freitas Neves Silma Regina Pereira |
| author_facet | Luiz Ricardo Goulart Bruna França Matias Colombo Mayara Ingrid Sousa Lima Maria Socorro A. de Andrade Juliana São Julião Adriana Freitas Neves Silma Regina Pereira |
| author_sort | Luiz Ricardo Goulart |
| collection | DOAJ |
| description | The role of the human papillomavirus (HPV) in the establishment of cervical cancer has driven studies to find more effective methods of viral detection so that early intervention strategies can be performed. However, the methods still have limitations, especially regarding detecting the different genotypes simultaneously. We have developed a high-throughput system using a single-tube nested-multiplex polymerase chain reaction (NMPCR) for the detection of 40 HPV genotypes using capillary electrophoresis. The NMPCR assay was compared to the Hybrid Capture 2 assay (HC2) with 40 women from the Northeast of Brazil (São Luis, MA), a high endemic region, where the HPV positivity was 75% and 37.5%, respectively. These results were validated by performing a molecular epidemiological study on 5223 Brazilian women undergoing gynecological examinations from 2009 to 2017, who presented with an HPV prevalence of 59%. Multiple infections were found in 62.5% and 58% of the patients from the endemic region and from the Brazilian women population, respectively, mostly presenting high-risk genotypes (90.5% and 60%, respectively). Considering cervical intraepithelial neoplasia and adenocarcinomas, the sensitivity and specificity were 97.5% and 100%, respectively. The NMPCR assay was also capable of identifying viral subtypes in cases of multiple infections, even with low viral loads (10<sup>−6</sup> ng/µL of HPV DNA). The NMPCR test is a promising and robust tool for HPV diagnostics and a screening tool for prevention of cervical cancer. |
| format | Article |
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| institution | OA Journals |
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| language | English |
| publishDate | 2024-11-01 |
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| series | Microorganisms |
| spelling | doaj-art-d42e46c46f1a4343ba95bbd99548d00c2025-08-20T02:05:06ZengMDPI AGMicroorganisms2076-26072024-11-011211232610.3390/microorganisms12112326Expanded HPV Genotyping by Single-Tube Nested-Multiplex PCR May Explain HPV-Related Disease RecurrenceLuiz Ricardo Goulart0Bruna França Matias Colombo1Mayara Ingrid Sousa Lima2Maria Socorro A. de Andrade3Juliana São Julião4Adriana Freitas Neves5Silma Regina Pereira6Institute of Biotechnology, Federal University of Uberlândia, Uberlândia 38400-902, MG, BrazilInstitute of Biotechnology, Federal University of Uberlândia, Uberlândia 38400-902, MG, BrazilLaboratory of Genetics and Molecular Biology, Department of Biology, Federal University of Maranhão, São Luís 65085-580, MA, BrazilHealth Department of the State of Maranhão, São Luís 65000-00, MA, BrazilBioGenetics Tecnologia Molecular Ltda., Uberlândia 38400-446, MG, BrazilInstitute of Biotechnology, Federal University of Catalão, Catalão 75705-220, GO, BrazilLaboratory of Genetics and Molecular Biology, Department of Biology, Federal University of Maranhão, São Luís 65085-580, MA, BrazilThe role of the human papillomavirus (HPV) in the establishment of cervical cancer has driven studies to find more effective methods of viral detection so that early intervention strategies can be performed. However, the methods still have limitations, especially regarding detecting the different genotypes simultaneously. We have developed a high-throughput system using a single-tube nested-multiplex polymerase chain reaction (NMPCR) for the detection of 40 HPV genotypes using capillary electrophoresis. The NMPCR assay was compared to the Hybrid Capture 2 assay (HC2) with 40 women from the Northeast of Brazil (São Luis, MA), a high endemic region, where the HPV positivity was 75% and 37.5%, respectively. These results were validated by performing a molecular epidemiological study on 5223 Brazilian women undergoing gynecological examinations from 2009 to 2017, who presented with an HPV prevalence of 59%. Multiple infections were found in 62.5% and 58% of the patients from the endemic region and from the Brazilian women population, respectively, mostly presenting high-risk genotypes (90.5% and 60%, respectively). Considering cervical intraepithelial neoplasia and adenocarcinomas, the sensitivity and specificity were 97.5% and 100%, respectively. The NMPCR assay was also capable of identifying viral subtypes in cases of multiple infections, even with low viral loads (10<sup>−6</sup> ng/µL of HPV DNA). The NMPCR test is a promising and robust tool for HPV diagnostics and a screening tool for prevention of cervical cancer.https://www.mdpi.com/2076-2607/12/11/2326HPV screeningcervical cancermolecular diagnosticsmolecular epidemiology |
| spellingShingle | Luiz Ricardo Goulart Bruna França Matias Colombo Mayara Ingrid Sousa Lima Maria Socorro A. de Andrade Juliana São Julião Adriana Freitas Neves Silma Regina Pereira Expanded HPV Genotyping by Single-Tube Nested-Multiplex PCR May Explain HPV-Related Disease Recurrence Microorganisms HPV screening cervical cancer molecular diagnostics molecular epidemiology |
| title | Expanded HPV Genotyping by Single-Tube Nested-Multiplex PCR May Explain HPV-Related Disease Recurrence |
| title_full | Expanded HPV Genotyping by Single-Tube Nested-Multiplex PCR May Explain HPV-Related Disease Recurrence |
| title_fullStr | Expanded HPV Genotyping by Single-Tube Nested-Multiplex PCR May Explain HPV-Related Disease Recurrence |
| title_full_unstemmed | Expanded HPV Genotyping by Single-Tube Nested-Multiplex PCR May Explain HPV-Related Disease Recurrence |
| title_short | Expanded HPV Genotyping by Single-Tube Nested-Multiplex PCR May Explain HPV-Related Disease Recurrence |
| title_sort | expanded hpv genotyping by single tube nested multiplex pcr may explain hpv related disease recurrence |
| topic | HPV screening cervical cancer molecular diagnostics molecular epidemiology |
| url | https://www.mdpi.com/2076-2607/12/11/2326 |
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