scholarly journals Detection of Human Papillomaviruses DNA in paired peripheral blood and cervix samples patients with cervical lesions and healthy individuals

2021 ◽  
Author(s):  
Kamylla Conceicao Gomes Nascimento ◽  
Elyda Goncalves Lima ◽  
Barbara Simas Chagas ◽  
Zhilbelly Mota Nunes ◽  
Marconi Rego Barros Junior ◽  
...  
Keyword(s):  
Peripheral Blood ◽  
Intraepithelial Neoplasia ◽  
Human Papillomaviruses ◽  
Healthy Individuals ◽  
Cervical Lesions ◽  
Hpv16 E6 ◽  
Hpv Dna ◽  
Sexually Transmitted ◽  
Standard Procedures ◽  
Hpv Types

This study evaluated the presence of HPV DNA in the cervix and peripheral blood of women with cervical intraepithelial neoplasia (CIN I, II, and III) and healthy individuals. Overall, 139 paired peripheral blood and cervix samples of healthy women and women with CIN I, II, and III (n= 68) were tested for HPV DNA by using standard procedures. PCR-sequencing determined HPV types. Quantification of HPV16 E6 and E2 genes was performed to determine viral load and physical state. HPV DNA was detected in the cervix (21.1% in healthy individuals; 48.8-55.5% in CIN patients), blood (46.4% in healthy individuals; 44.1-77.7% in CIN patients), and paired peripheral blood and cervix samples (24% in healthy individuals; 32.5-44.4% in CIN patients). The most frequent types found in the cervix were HPV16, 18, 31, 33, 58, and 70, while HPV16, 18, 33, 58, and 66 were the most frequent types found in the blood. HPV DNA in the cervix was associated with previous sexually transmitted infections (STIs) (P=0.023; OR: 2.978; CI:1.34-7.821), HPV DNA in the blood (P=0.000; OR: 3.369; CI:3.700-18.540), and cervical lesions (CIN I/II or III) (P=0.001; OR: 3.369; CI:1.634-6.945). Binomial Logistic regression showed that HPV DNA in the blood (P=0.000; OR: 9.324; CI:3.612-24.072) and cervical lesions (P=0.011; OR: 3.622; CI:1.338-9.806) were associated with HPV DNA in the cervix. However, we did not find an association between HPV DNA in blood and cervical lesions (P=0.385). Our results showed that, although there is an association between HPV DNA in the cervix with HPV DNA in blood, only HPV DNA found in the cervix was associated with cervical lesions.

scholarly journals Detection of Human Papillomavirus DNA in Paired Peripheral Blood and Cervix Samples in Patients with Cervical Lesions and Healthy Individuals

2021 ◽  
Vol 10 (21) ◽  
pp. 5209
Author(s):  
Kamylla Conceição Gomes do Nascimento ◽  
Élyda Gonçalves de Lima ◽  
Zhilbelly da Mota Nunes ◽  
Marconi Rêgo Barros ◽  
Marcus Vinícius de Aragão Batista ◽  
...  
Keyword(s):  
Human Papillomavirus ◽  
Peripheral Blood ◽  
Intraepithelial Neoplasia ◽  
Healthy Individuals ◽  
Cervical Lesions ◽  
Hpv16 E6 ◽  
Hpv Dna ◽  
Sexually Transmitted ◽  
Standard Procedures ◽  
Papillomavirus Dna

This study evaluated the presence of Human Papillomavirus (HPV) DNA in the cervix and peripheral blood of women with cervical intraepithelial neoplasia (CIN I, II, and III) and healthy individuals. Overall, 139 paired peripheral blood and cervix samples of healthy women and women with CIN I, II, and III (n = 68) were tested for HPV DNA by using standard procedures. Polymerase chain reaction (PCR) sequencing determined HPV types. Quantification of HPV16 E6 and E2 genes was performed to determine viral load and physical state. HPV DNA was detected in the cervix (21.1% in healthy individuals; 48.8–55.5% in CIN patients), blood (46.4% in healthy individuals; 44.1–77.7% in CIN patients) and paired peripheral blood and cervix samples (24% in healthy individuals; 32.5–44.4% in CIN patients). The most frequent types found in the cervix were HPV16, 18, 31, 33, 58, and 70, while HPV16, 18, 33, 58, and 66 were the most frequent types found in the blood. HPV DNA in the cervix was associated with previous sexually transmitted infections (STIs) (p = 0.023; OR: 2.978; CI:1.34–7.821), HPV DNA in the blood (p = 0.000; OR: 8.283; CI:3.700–18.540), and cervical lesions (CIN I/II or III) (p = 0.007). Binomial logistic regression showed that HPV DNA in the blood (p = 0.000; OR: 9.324; CI:3.612–24.072) and cervical lesions (p = 0.011; OR: 3.622; CI:1.338–9.806) were associated with HPV DNA in the cervix. However, we did not find an association between HPV DNA in the blood and cervical lesions (p = 0.385). Our results showed that only HPV DNA found in the cervix was associated with cervical lesions.

scholarly journals The Epidemiology of Human Papillomavirus Infection and Cervical Cancer

2007 ◽  
Vol 23 (4) ◽  
pp. 213-227 ◽  
Author(s):  
F. Xavier Bosch ◽  
Silvia de Sanjosé
Keyword(s):  
Cervical Cancer ◽  
Invasive Cervical Cancer ◽  
Intraepithelial Neoplasia ◽  
Hpv Infection ◽  
Human Papillomaviruses ◽  
Transmitted Infection ◽  
Hpv Dna ◽  
Sexually Transmitted ◽  
Papillomavirus Infection ◽  
Simplex Virus

Cervical cancer has been recognized as a rare outcome of a common Sexually Transmitted Infection (STI). The etiologic association is restricted to a limited number of viral types of the family of the Human Papillomaviruses (HPVs). The association is causal in nature and under optimal testing systems, HPV DNA can be identified in all specimens of invasive cervical cancer. As a consequence, it has been claimed that HPV infection is a necessary cause of cervical cancer. The evidence is consistent worldwide and implies both the Squamous Cell Carcinomas (SCC), the adenocarcinomas and the vast majority (i.e. > 95%) of the immediate precursors, namely High Grade Squamous Intraepithelial Lesions (HSIL)/Cervical Intraepithelial Neoplasia 3 (CIN3)/Carcinomain situ. Co-factors that modify the risk among HPV DNA positive women include the use of oral contraceptives (OC) for five or more years, smoking, high parity (five or more full term pregnancies) and previous exposure to other sexually transmitted diseases such as Chlamydia Trachomatis (CT) and Herpes Simplex Virus type 2 (HSV-2). Women exposed to the Human Immunodeficiency Virus (HIV) are at high risk for HPV infection, HPV DNA persistency and progression of HPV lesions to cervical cancer.

scholarly journals Phylogeny and polymorphism in the E6 and E7 of human papillomavirus: alpha-9 (HPV16, 31, 33, 52, 58), alpha-5 (HPV51), alpha-6 (HPV53, 66), alpha-7 (HPV18, 39, 59, 68) and alpha-10 (HPV6, 44) in women from Shanghai

2019 ◽  
Vol 14 (1) ◽  
Author(s):  
Junwei Zhao ◽  
Qin Zhan ◽  
Junhan Guo ◽  
Min Liu ◽  
Yetian Ruan ◽  
...  
Keyword(s):  
Phylogenetic Trees ◽  
Intraepithelial Neoplasia ◽  
Human Papillomaviruses ◽  
Therapeutic Interventions ◽  
Cervical Lesions ◽  
Intraepithelial Lesion ◽  
Species Groups ◽  
Hpv Types ◽  
E6 And E7 ◽  
Alpha 7

Abstract Background Persistent infection with human papillomaviruses (HPVs) has been associated with cervical intraepithelial neoplasia (CIN) and cervical cancer. However, why only a fraction of HPV cases progress to cancer is still unclear. Methods We focused on the heterogeneity, classification, evolution and dispersal of variants for 14 common HPV types in 262 HPV-positive patients with cervical lesions. The E6 and E7 genes of HPV were sequenced and compared with the HPV reference for sequence analysis. Phylogenetic trees were constructed using the neighbour-joining tree method with MEGA 7.0. Results In this study, 233 E6 and 212 E7 sequences were successfully amplified by PCR, and these sequences were divided into 5 species groups: alpha-9 (HPV16, 31, 33, 52, 58), alpha-5 (HPV51), alpha-6 (HPV53, 66), alpha-7 (HPV18, 39, 59, 68) and alpha-10 (HPV6, 44). The incidence of high-grade squamous intraepithelial lesion (HSIL) in patients infected with alpha-9 HPV was significantly increased compared with other groups (P < 0.0001), especially HPV16 (P < 0.0001). Strikingly, E7 had significantly fewer nonsynonymous variants in the HSIL compared to <HSIL groups (P = 3.17× 10− 4). The A388C (K93 N) variation in HPV58 E6 can significantly reduce the risk of HSIL (P = 0.015). However, T7220G (D32E) variation in HPV16 E6 and A7689G (N29S) in HPV16 E7 increased the incidence of HSIL compared to the <HSIL group (P = 0.036 and 0.022). Conclusions Strict conservation of E7 is important for HPV carcinogenicity, especially N29 of HPV16. The findings in this work provide preventative/therapeutic interventions for HPV infections and CIN.

scholarly journals Quantification of HPV16 E6/E7 mRNA Spliced Isoforms Viral Load as a Novel Diagnostic Tool for Improving Cervical Cancer Screening

2018 ◽  
Vol 7 (12) ◽  
pp. 530 ◽  
Author(s):  
Claire Camus ◽  
Sébastien Vitale ◽  
Céline Loubatier ◽  
Guillaume Pénaranda ◽  
Hacène Khiri ◽  
...  
Keyword(s):  
Cervical Cancer ◽  
Cancer Screening ◽  
Cervical Intraepithelial Neoplasia ◽  
Cervical Cancer Screening ◽  
Diagnostic Performance ◽  
Pap Test ◽  
Intraepithelial Neoplasia ◽  
Human Papillomaviruses ◽  
Cervical Lesions ◽  
Hpv16 E6

High-risk human papillomaviruses (HPVs) have been identified as the main contributors to cervical cancer. Despite various diagnostic tools available, including the predominant Papanicolaou test (Pap test), technical limitations affect the efficiency of cervical cancer screening. The aim of this study was to evaluate the diagnostic performance of spliced HPV16 E6/E7 mRNA viral loads (VL) for grade 2 or higher cervical intraepithelial neoplasia diagnosis. A new dedicated (quantitative reverse transcription polymerase chain reaction) qRT-PCR assay was developed, allowing selective quantification of several HPV16 E6/E7 mRNA: Full length (FL) with or without all or selected spliced forms (total E6/E7 mRNA corresponding to SP + E6^E7 mRNA (T), + spliced E6/E7 mRNA containing intact E7 ORF (SP), and E6/E7 mRNA containing disrupted E6 and E7 ORFs calculated by the following subtraction T-SP (E6^E7)). Twenty HPV16 DNA and mRNA positive uterine cervical smears representative of all cytological and histological stages of severity were tested. We have shown that all E6/E7 mRNA isoforms expression levels were significantly increased in high grade cervical lesions. Statistical analysis demonstrated that the SP-E6/E7 VL assay exhibited: (i) The best diagnostic performance for identification of both cervical intraepithelial neoplasia (CIN)2+ (90% (56–100) sensitivity and specificity) and CIN3+ (100% (72–100) sensitivity and 79% (49–95) specificity) lesions; (ii) a greater sensitivity compared to the Pap test for CIN2+ lesions detection (80% (44–97)); (iii) a predictive value of the histological grade of cervical lesions in 67% of atypical squamous cells of unknown significance (ASC-US) and 100% of low-grade (LSIL) patients. Overall, these results highlight the value of SP-E6/E7 mRNA VL as an innovative tool for improving cervical cancer screening.

Human papillomavirus (HPV) study of 2916 cytological samples by PCR and DNA sequencing: genotype spectrum of patients from the west German area

2004 ◽  
Vol 53 (2) ◽  
pp. 125-128 ◽  
Author(s):  
Norbert Speich ◽  
Christoph Schmitt ◽  
Reinhard Bollmann ◽  
Magdolna Bollmann
Keyword(s):  
Direct Sequencing ◽  
Intraepithelial Neoplasia ◽  
Human Papillomaviruses ◽  
Tissue Samples ◽  
Hpv Dna ◽  
Pcr Products ◽  
Hpv Types ◽  
Spectrum Of Patients ◽  
Consensus Primers

Human papillomaviruses (HPVs) are aetiological agents for cervical cancer. More than 70 different HPV types that infect genital mucosa have been found. In order to develop a sensitive and specific detection and typing assay, a PCR/direct sequencing approach was used. Two pairs of consensus primers were used for amplification of HPV DNA and the PCR products obtained were analysed by automated sequencing. Sequences were compared with those in GenBank by using the blast program. In this study, 2916 cytological samples were screened for HPV, as well as for triage. Nine hundred and forty-eight (32.5 %) samples were positive for HPV, of which 134 harboured more than one HPV type. Of the 948 PCR-positive samples, 648 were typed. Thirty-nine different HPV types were identified by sequencing. The two most frequently found HPV types, 16 and 31, together accounted for 36.3 % of the sequences (26.2 and 10.1 %, respectively). This group was followed by HPV types 6 (5.7 %), 18 (5.3 %), 58 (4.5 %), 61 (4.5 %), 53 (4.4 %), 42 (4.3 %) and 51 (4.0 %). All other types were detected at frequencies <4 % and eight types were detected only once. PCR/direct sequencing is a reliable method for routine detection of HPV in cytological samples. The data presented here suggest a complex distribution of HPV types in the population tested. The results accentuate the importance of PCR-based techniques in HPV diagnosis, as hybridization-based methods can only detect a limited number of infections. This method can also be applied easily to the analysis of tissue samples and it therefore also allows type-specific follow-up of women who have been treated for cervical intraepithelial neoplasia.

scholarly journals Prevalence and type distribution of human papillomavirus infection among women with different degrees of cervical cytological abnormalities in Sicily (Italy)

2016 ◽  
Vol 31 (2) ◽  
Author(s):  
Concetta Franchina ◽  
Carmela M. Costanzo ◽  
Raffaela Russo ◽  
Concetta I. Palermo ◽  
Guido Scalia
Keyword(s):  
Pap Smear ◽  
Blot Hybridization ◽  
Human Papillomaviruses ◽  
Multiple Infections ◽  
Low Grade ◽  
Genotype Distribution ◽  
Cervical Lesions ◽  
Hpv Dna ◽  
Papillomavirus Infection ◽  
Hpv Genotypes

Human papillomaviruses (HPVs) are etiological agents of cervical cancer. In the absence of Pap smear alterations, high-risk HPV DNA can be detected in cervical samples. The prevalence of papillomavirus infection and their genotype distribution varies greatly across populations. The aims of this study were: i) to assess the prevalences of HPV genotypes in people living in Eastern Sicily (Italy) and the frequency of HPV multiple infections; ii) to evaluate the association between HPV genotypes and cervical lesions in order to improve the epidemiological knowledge useful for monitoring or treating infected women. Nested PCR and reverse dot/blot hybridization were used for the detection and typing of HPV DNA in 315 women who had had an abnormal PAP-smear. HPV DNA test was positive in 70.5% cases; the prevalence was 50% in atypical squamous cells of undetermined significance (ASCUS), 80.8% in low grade-, and 76.2% in high grade-squamous intraepithelial lesion (H-SIL). The genotype distribution showed a predominance of HPV-16 (56.7%) followed by HPV-18 (12.2%), HPV-31 (9.5%) and HPV-6 (9.5%). Multiple infections were detected in 35.1% of the infected patients. High frequency of positive results for HPV was confirmed and, even in case of ASCUS, patients should be taken into account for genotyping. Our data indicate that multiple infections are consistent in women with low-grade lesions while they are less frequent in women with H-SIL. This could reinforce the theory of the multi-stage cancer model, by which one HPV type becomes predominant along with the progression of cervical lesion severity.

Comparison of Human Papillomavirus Detection and Typing by Hybrid Capture 2, Linear Array, DNA Chip, and Cycle Sequencing in Cervical Swab Samples

2009 ◽  
Vol 19 (2) ◽  
pp. 266-272 ◽  
Author(s):  
Jae Kwan Lee ◽  
Mi Kyung Kim ◽  
Seung Hun Song ◽  
Jin Hwa Hong ◽  
Kyung Jin Min ◽  
...  
Keyword(s):  
High Risk ◽  
Linear Array ◽  
Intraepithelial Neoplasia ◽  
Dna Chip ◽  
Substantial Agreement ◽  
Cervical Lesions ◽  
Cycle Sequencing ◽  
Hpv Dna ◽  
Hpv Dna Tests ◽  
High Risk Hpv

Although the Hybrid Capture II (HC II) assay can detect 13 high-risk human papillomavirus (HPVs), it does not yield any genotype-specific information. We evaluated the performance of 4 HPV DNA tests, namely, HC II, Linear Array (LA), DNA chip, and cycle sequencing for their capacity to detect the presence of high-risk HPV DNA and HPV-associated cervical lesions. Seventy-six women who were referred to the colposcopy clinic for abnormal cytology were enrolled. The women were examined using liquid-based cytology, colposcopy-directed biopsy, and HPV DNA tests. After DNA extraction from a single sample, HPV DNA tests were performed by all 4 methods on the same specimen. The LA test has higher HPV-positive rates than HC II for cervical intraepithelial neoplasia I (83.3% vs 61.1%;P< 0.01) and for cervical intraepithelial neoplasia II and more severe lesions (100.0% vs 80.0%;P< 0.01). The concordance between the DNA chip and LA tests was 89.5%, confirming substantial agreement (κcoefficient = 0.73), and the concordance between HC II and the DNA chip was 80.3%, also showing substantial agreement (κcoefficient = 0.738). The concordance for 15 high-risk HPV genotypes between LA and sequencing was 82.5% with aκvalue of 0.536. Furthermore, the LA test was more sensitive in the detection of high-grade cervical lesions than HC II (100% vs 92.3%,P< 0.01). The LA test showed superior sensitivity in the detection of clinically relevant HPV infections and has proven to be an accurate tool for identifying individual HPV types, especially in cases of multiple HPV infections.

scholarly journals Human papillomavirus and Its Association with Other Sexually Transmitted Coinfection among Sexually Active Women from the Northeast of Brazil

2020 ◽  
Vol 2020 ◽  
pp. 1-8
Author(s):  
Ana Paula Almeida Cunha ◽  
Ilka Kassandra Pereira Belfort ◽  
Francisco Pedro Belfort Mendes ◽  
Gerusinete Rodrigues Bastos dos Santos ◽  
Lucas Henrique de Lima Costa ◽  
...  
Keyword(s):  
Chlamydia Trachomatis ◽  
Neisseria Gonorrhoeae ◽  
Trichomonas Vaginalis ◽  
Positive Association ◽  
Hpv Infection ◽  
Cervical Lesions ◽  
Hpv Dna ◽  
Sexually Transmitted ◽  
Epithelial Lesions ◽  
Intraepithelial Lesions

Objective. To verify the association between HPV infection and the presence of coinfections (Chlamydia trachomatis, Trichomonas vaginalis, and Neisseria gonorrhoeae) in women in the state of Maranhão. Methods. HPV-DNA detection was performed by the nested PCR, using the primers PGMY09/11 and GP + 5/GP + 6. For the identification of sexually transmitted agents, conventional PCR was performed using the following primers: KL1/KL2 (Chlamydia trachomatis), TVA5/TVA6 (Trichomonas vaginalis), and HO1/HO3 (Neisseria gonorrhoeae). DNA-HPV positive samples were subjected to automated sequencing for genotyping. Results. Among the 353 women evaluated, 204 (57.8%) had HPV-DNA, of which 140 (68.6%) exhibited HPV/STIs, while 64 (31.4%) had the only HPV. T. vaginalis infection showed a positive association with HPV ( p = 0.003 ). Women without cervical lesions were predominant (327/92.6%); however, the largest number of lesions was reported in women who had HPV/coinfections (18/8.8%). Multiple regression analysis showed that both HPV only and the concomitant presence of HPV/STI were able to indicate the occurrence of epithelial lesions (R = 0.164; R2 = 0.027). Conclusion. The findings suggest that the presence of T. vaginalis can contribute to HPV infection, and HPV/IST association may influence the development of cervical intraepithelial lesions that are precursors of cervical cancer.

Discovery of candidate gene expression signatures in peripheral blood for the screening of cervical cancer

2020 ◽  
Vol 14 (2) ◽  
pp. 109-118 ◽  
Author(s):  
Qiuling Ma ◽  
Yong Shao ◽  
Wei Chen ◽  
Cheng Quan ◽  
Yanhui Zhu ◽  
...  
Keyword(s):  
Gene Expression ◽  
Cervical Cancer ◽  
Real Time ◽  
Real Time Pcr ◽  
Peripheral Blood ◽  
Area Under The Curve ◽  
Intraepithelial Neoplasia ◽  
Sequencing Analysis ◽  
Quantitative Real Time Pcr ◽  
Cervical Lesions

Aim: To investigate whether cervical cancer (CC) and cervical intraepithelial neoplasia (CIN) can be screened by analyzing gene expression profiling of peripheral blood. Methods: RNA-sequencing analysis of blood was performed on 11 CC patients, 21 CIN patients and 19 healthy controls (H). Fifty-nine genes were validated by quantitative real-time PCR using blood samples from 46 H, 83 CC and 32 CIN patients. Results: There were significant differences in the expression levels of six genes between CC and H, five genes between CIN and H and four genes between CC and CIN (p < 0.05). Four genes discriminated cervical lesions from H with a sensitivity of 82.61%, a specificity of 87.83% and an area under the curve of 0.8981. Three genes discriminated CC from CIN with a sensitivity of 53.13%, a specificity of 96.39% and an area under the curve of 0.7786. Conclusion: Our findings provided a promising noninvasive quantitative real-time PCR diagnostic assay of CC and CIN.

scholarly journals Virus-like Particle-Based L2 Vaccines against HPVs: Where Are We Today?

Viruses ◽  
2019 ◽  
Vol 12 (1) ◽  
pp. 18 ◽  
Author(s):  
Rashi Yadav ◽  
Lukai Zhai ◽  
Ebenezer Tumban
Keyword(s):  
Amino Acids ◽  
Capsid Protein ◽  
Hpv Vaccine ◽  
Genital Warts ◽  
Human Papillomaviruses ◽  
Target Antigen ◽  
Sexually Transmitted ◽  
Virus Like Particle ◽  
Protective Antibodies ◽  
Hpv Types

Human papillomaviruses (HPVs) are the most common sexually transmitted infections worldwide. Ninety percent of infected individuals clear the infection within two years; however, in the remaining 10% of infected individuals, the infection(s) persists and ultimately leads to cancers (anogenital cancers and head and neck cancers) and genital warts. Fortunately, three prophylactic vaccines have been approved to protect against HPV infections. The most recent HPV vaccine, Gardasil-9 (a nonavalent vaccine), protects against seven HPV types associated with ~90% of cervical cancer and against two HPV types associated with ~90% genital warts with little cross-protection against non-vaccine HPV types. The current vaccines are based on virus-like particles (VLPs) derived from the major capsid protein, L1. The L1 protein is not conserved among HPV types. The minor capsid protein, L2, on the other hand, is highly conserved among HPV types and has been an alternative target antigen, for over two decades, to develop a broadly protective HPV vaccine. The L2 protein, unlike the L1, cannot form VLPs and as such, it is less immunogenic. This review summarizes current studies aimed at developing HPV L2 vaccines by multivalently displaying L2 peptides on VLPs derived from bacteriophages and eukaryotic viruses. Recent data show that a monovalent HPV L1 VLP as well as bivalent MS2 VLPs displaying HPV L2 peptides (representing amino acids 17–36 and/or consensus amino acids 69–86) elicit robust broadly protective antibodies against diverse HPV types (6/11/16/18/26/31/33/34/35/39/43/44/45/51/52/53/56/58/59/66/68/73) associated with cancers and genital warts. Thus, VLP-based L2 vaccines look promising and may be favorable, in the near future, over current L1-based HPV vaccines and should be explored further.

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