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A case of HPV-53-related cervical cancer in an elderly patient.

Lieveld M, Padalko E, Praet M, Vanden Broeck D - Clin Interv Aging (2014)

View Article: PubMed Central - PubMed

Affiliation: Department of Uro/gynaecology, Ghent University Hospital, Ghent, Belgium.

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It is suggested that HPV53 is present exclusively in the cervical cancer cells, lymph node metastases, and atypical urinary cells of one single case while the surrounding CIN2+ tissue revealed ten different HPV strains... Unfortunately, the HPV genotype results for lymph nodes and urinary cells are not presented while these results underline the potential role of HPV53 in oncogenesis... In Figure 3 in Zappacosta et al two melting peaks (-d(RFU)/dt) for HPV53 are displayed, one detected by probe A in plasmid DNA (referred to as a positive internal control), and one detected by probe B in cervical cancer cells... The internal positive control used in Anyplex II HPV28 is human β-globin and not plasmid DNA as indicated by Zappacosta et al... It is included to monitor whether the sample is adequate for analysis and to exclude polymerase chain reaction (PCR)-inhibition which may lead to false negative results... Therefore, it is expected that HPV53 infection in cervical cancer cells (right melting curve in Figure 3, Zappacosta et al) will give the exact same melting profile as HPV53 observed in the positive control (left melting curve in Figure 3, Zappacosta et al)... The Anyplex II HPV28 is based on melting curve analysis using the TOCE™ technology... To our knowledge, HC2 HPV test has not yet been optimized for urine samples., In-house optimization of urine samples for HC2 HPV could have been performed, but this is not specified in the article... It is also unclear why the authors carried out the HC2 HPV test since the results generated by Anyplex II HPV28 and HC2 are not compared with each other in the Results section... However, it is unclear whether a negative control was included in the experiments in order to exclude cross-contamination... Validation for cross-contamination could be performed beforehand, but this is not reported in the article... In conclusion, based on the observation that the melting curve of the HPV type detected in the cervical cancer cells does not correspond with the melting curve for HPV53 plasmid DNA, it can be deduced that another HPV type might be involved in the cervical cancer cells... Hence, it cannot be concluded that HPV53 is solely responsible in oncogenesis in this case as previously suggested by Zappacosta et al.

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Real-time PCR assay performed on tissues (invasive cervical cancer, CIN2+ lesion adjacent to invasive neoplasia, metastatic lymph nodes, non-metastatic lymph nodes, and thyroid), and on liquid-based urinary samples.Notes: Anyplex II HPV28 detection assay simultaneously detects 28 HPV genotypes. β-globin gene is used as a positive internal control to identify processed specimens containing substances that may interfere with PCR amplification. Thyroid tissue was used as an external HPV-negative control. (A) Invasive cervical cancer showing positive result for HPV-53. (B) CIN2+ lesion, in which positive results for HPVs 16, 35, 39, 40, 53, 54, 59, 61, 68 and 82 have been found. (C) Metastatic lymph nodes, showing HPV-53 positive result. (D) Non-metastatic lymph nodes, testing as HPV-negative. (E) Thyroid tissue testing as HPV-negative. (F) Urinary samples, in which HPV-53 has been found. (G) Melting curve of HPV-53 control included in the PC3 mix of set B, demonstrating the melting curve shape and Tm. The melting profile of samples in which HPV-53 has been found was shown to have a Tm of 76.5°C.Abbreviations: PCR, polymerase chain reaction; CIN2+, cervical intraepithelial neoplasia grade 2-or-worse; -d(RFU/dT), change in rate of fluorescence units against temperature; IC, internal control; PC3 of set B, positive control of set B, consisting of a mix of positive controls, including those of HPVs 73, 53, 70, 6, and 11; Tm, melting temperature; HPV, human papillomavirus.
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f1-cia-9-1933: Real-time PCR assay performed on tissues (invasive cervical cancer, CIN2+ lesion adjacent to invasive neoplasia, metastatic lymph nodes, non-metastatic lymph nodes, and thyroid), and on liquid-based urinary samples.Notes: Anyplex II HPV28 detection assay simultaneously detects 28 HPV genotypes. β-globin gene is used as a positive internal control to identify processed specimens containing substances that may interfere with PCR amplification. Thyroid tissue was used as an external HPV-negative control. (A) Invasive cervical cancer showing positive result for HPV-53. (B) CIN2+ lesion, in which positive results for HPVs 16, 35, 39, 40, 53, 54, 59, 61, 68 and 82 have been found. (C) Metastatic lymph nodes, showing HPV-53 positive result. (D) Non-metastatic lymph nodes, testing as HPV-negative. (E) Thyroid tissue testing as HPV-negative. (F) Urinary samples, in which HPV-53 has been found. (G) Melting curve of HPV-53 control included in the PC3 mix of set B, demonstrating the melting curve shape and Tm. The melting profile of samples in which HPV-53 has been found was shown to have a Tm of 76.5°C.Abbreviations: PCR, polymerase chain reaction; CIN2+, cervical intraepithelial neoplasia grade 2-or-worse; -d(RFU/dT), change in rate of fluorescence units against temperature; IC, internal control; PC3 of set B, positive control of set B, consisting of a mix of positive controls, including those of HPVs 73, 53, 70, 6, and 11; Tm, melting temperature; HPV, human papillomavirus.

Mentions: In Figure 3 in Zappacosta et al1 two melting peaks (-d(RFU)/dt) for HPV53 are displayed, one detected by probe A in plasmid DNA (referred to as a positive internal control), and one detected by probe B in cervical cancer cells. However, Anyplex II HPV28 only uses one target specific primer/catcher/pitcher set for each HPV type. The internal positive control used in Anyplex II HPV28 is human β-globin and not plasmid DNA as indicated by Zappacosta et al.1 It is included to monitor whether the sample is adequate for analysis and to exclude polymerase chain reaction (PCR)-inhibition which may lead to false negative results. The test also includes external positive controls which consist of HPV type-specific plasmids, in order to validate whether the prepared master mix accurately detects the indicated HPV types. Nevertheless, the authors could opt to use synthesized plasmid HPV53 DNA as an extra internal positive control by adding it to the sample but this could not be derived from the article. In either case, melting curve analysis of the positive control will give a melting profile consisting of melting curves specific for each HPV type included in the positive control (personal communication Seegene). Because the shape of the melting curve and the position of the peak is a signature for each HPV type, infection with a particular HPV type will give rise to a particular melting curve. Therefore, it is expected that HPV53 infection in cervical cancer cells (right melting curve in Figure 3, Zappacosta et al1) will give the exact same melting profile as HPV53 observed in the positive control (left melting curve in Figure 3, Zappacosta et al1). This is not the case, as it can be deduced from Figure 3, Zappacosta et al. The melting peak of HPV53 in the positive control differs approximately 8°C from the HPV type detected in cervical cancer cells.


A case of HPV-53-related cervical cancer in an elderly patient.

Lieveld M, Padalko E, Praet M, Vanden Broeck D - Clin Interv Aging (2014)

Real-time PCR assay performed on tissues (invasive cervical cancer, CIN2+ lesion adjacent to invasive neoplasia, metastatic lymph nodes, non-metastatic lymph nodes, and thyroid), and on liquid-based urinary samples.Notes: Anyplex II HPV28 detection assay simultaneously detects 28 HPV genotypes. β-globin gene is used as a positive internal control to identify processed specimens containing substances that may interfere with PCR amplification. Thyroid tissue was used as an external HPV-negative control. (A) Invasive cervical cancer showing positive result for HPV-53. (B) CIN2+ lesion, in which positive results for HPVs 16, 35, 39, 40, 53, 54, 59, 61, 68 and 82 have been found. (C) Metastatic lymph nodes, showing HPV-53 positive result. (D) Non-metastatic lymph nodes, testing as HPV-negative. (E) Thyroid tissue testing as HPV-negative. (F) Urinary samples, in which HPV-53 has been found. (G) Melting curve of HPV-53 control included in the PC3 mix of set B, demonstrating the melting curve shape and Tm. The melting profile of samples in which HPV-53 has been found was shown to have a Tm of 76.5°C.Abbreviations: PCR, polymerase chain reaction; CIN2+, cervical intraepithelial neoplasia grade 2-or-worse; -d(RFU/dT), change in rate of fluorescence units against temperature; IC, internal control; PC3 of set B, positive control of set B, consisting of a mix of positive controls, including those of HPVs 73, 53, 70, 6, and 11; Tm, melting temperature; HPV, human papillomavirus.
© Copyright Policy
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC4234393&req=5

f1-cia-9-1933: Real-time PCR assay performed on tissues (invasive cervical cancer, CIN2+ lesion adjacent to invasive neoplasia, metastatic lymph nodes, non-metastatic lymph nodes, and thyroid), and on liquid-based urinary samples.Notes: Anyplex II HPV28 detection assay simultaneously detects 28 HPV genotypes. β-globin gene is used as a positive internal control to identify processed specimens containing substances that may interfere with PCR amplification. Thyroid tissue was used as an external HPV-negative control. (A) Invasive cervical cancer showing positive result for HPV-53. (B) CIN2+ lesion, in which positive results for HPVs 16, 35, 39, 40, 53, 54, 59, 61, 68 and 82 have been found. (C) Metastatic lymph nodes, showing HPV-53 positive result. (D) Non-metastatic lymph nodes, testing as HPV-negative. (E) Thyroid tissue testing as HPV-negative. (F) Urinary samples, in which HPV-53 has been found. (G) Melting curve of HPV-53 control included in the PC3 mix of set B, demonstrating the melting curve shape and Tm. The melting profile of samples in which HPV-53 has been found was shown to have a Tm of 76.5°C.Abbreviations: PCR, polymerase chain reaction; CIN2+, cervical intraepithelial neoplasia grade 2-or-worse; -d(RFU/dT), change in rate of fluorescence units against temperature; IC, internal control; PC3 of set B, positive control of set B, consisting of a mix of positive controls, including those of HPVs 73, 53, 70, 6, and 11; Tm, melting temperature; HPV, human papillomavirus.
Mentions: In Figure 3 in Zappacosta et al1 two melting peaks (-d(RFU)/dt) for HPV53 are displayed, one detected by probe A in plasmid DNA (referred to as a positive internal control), and one detected by probe B in cervical cancer cells. However, Anyplex II HPV28 only uses one target specific primer/catcher/pitcher set for each HPV type. The internal positive control used in Anyplex II HPV28 is human β-globin and not plasmid DNA as indicated by Zappacosta et al.1 It is included to monitor whether the sample is adequate for analysis and to exclude polymerase chain reaction (PCR)-inhibition which may lead to false negative results. The test also includes external positive controls which consist of HPV type-specific plasmids, in order to validate whether the prepared master mix accurately detects the indicated HPV types. Nevertheless, the authors could opt to use synthesized plasmid HPV53 DNA as an extra internal positive control by adding it to the sample but this could not be derived from the article. In either case, melting curve analysis of the positive control will give a melting profile consisting of melting curves specific for each HPV type included in the positive control (personal communication Seegene). Because the shape of the melting curve and the position of the peak is a signature for each HPV type, infection with a particular HPV type will give rise to a particular melting curve. Therefore, it is expected that HPV53 infection in cervical cancer cells (right melting curve in Figure 3, Zappacosta et al1) will give the exact same melting profile as HPV53 observed in the positive control (left melting curve in Figure 3, Zappacosta et al1). This is not the case, as it can be deduced from Figure 3, Zappacosta et al. The melting peak of HPV53 in the positive control differs approximately 8°C from the HPV type detected in cervical cancer cells.

View Article: PubMed Central - PubMed

Affiliation: Department of Uro/gynaecology, Ghent University Hospital, Ghent, Belgium.

AUTOMATICALLY GENERATED EXCERPT
Please rate it.

It is suggested that HPV53 is present exclusively in the cervical cancer cells, lymph node metastases, and atypical urinary cells of one single case while the surrounding CIN2+ tissue revealed ten different HPV strains... Unfortunately, the HPV genotype results for lymph nodes and urinary cells are not presented while these results underline the potential role of HPV53 in oncogenesis... In Figure 3 in Zappacosta et al two melting peaks (-d(RFU)/dt) for HPV53 are displayed, one detected by probe A in plasmid DNA (referred to as a positive internal control), and one detected by probe B in cervical cancer cells... The internal positive control used in Anyplex II HPV28 is human β-globin and not plasmid DNA as indicated by Zappacosta et al... It is included to monitor whether the sample is adequate for analysis and to exclude polymerase chain reaction (PCR)-inhibition which may lead to false negative results... Therefore, it is expected that HPV53 infection in cervical cancer cells (right melting curve in Figure 3, Zappacosta et al) will give the exact same melting profile as HPV53 observed in the positive control (left melting curve in Figure 3, Zappacosta et al)... The Anyplex II HPV28 is based on melting curve analysis using the TOCE™ technology... To our knowledge, HC2 HPV test has not yet been optimized for urine samples., In-house optimization of urine samples for HC2 HPV could have been performed, but this is not specified in the article... It is also unclear why the authors carried out the HC2 HPV test since the results generated by Anyplex II HPV28 and HC2 are not compared with each other in the Results section... However, it is unclear whether a negative control was included in the experiments in order to exclude cross-contamination... Validation for cross-contamination could be performed beforehand, but this is not reported in the article... In conclusion, based on the observation that the melting curve of the HPV type detected in the cervical cancer cells does not correspond with the melting curve for HPV53 plasmid DNA, it can be deduced that another HPV type might be involved in the cervical cancer cells... Hence, it cannot be concluded that HPV53 is solely responsible in oncogenesis in this case as previously suggested by Zappacosta et al.

Show MeSH
Related in: MedlinePlus