scholarly journals Electrochemical DNA biosensor for the multianalyte detection of HPV-16 and HPV-18 in oral and cervical lesions

Author(s):  
Thanyarat Chaibun ◽  
Patcharanin Thanasapburachot ◽  
Patutong Chatchawal ◽  
Su Yin Lee ◽  
Sirimanas Jiaranuchart ◽  
...  

Abstract Infection with high-risk human papillomavirus (HPV) is a major risk factor for oral and cervical cancers. In this study, we developed an electrochemical DNA biosensor for detection of HPV-16 and HPV-18, which are the 2 most prevalent genotypes for development of oral and cervical cancers. The assay involves the sandwich hybridization of the HPV target to silica-redox dye reporter probe and capture probe, followed by electrochemical detection. The sensor was found to be highly specific and sensitive, with detection limit of 22 fM for HPV-16 and 20 fM for HPV-18, between the range of 1 fM to 1 µM. Evaluation with oral and cervical samples showed that the biosensor result was consistent with the nested PCR /gel electrophoresis detection. The biosensor assay could be completed within 90 minutes. Due to its simplicity, rapidity and high sensitivity, this biosensor could be used as an alternative method for HPV detection in clinical laboratories. [151 words]

2021 ◽  
Vol 15 (09) ◽  
pp. 1339-1345
Author(s):  
Richard Tagne Simo ◽  
Arsène G Djoko Nono ◽  
Hervet Paulin Fogang Dongmo ◽  
Paul F Seke Etet ◽  
Bertrand Kiafon Fonyuy ◽  
...  

Introduction: Various Human papillomavirus (HPV) types cause cervical cancer, and represent the primary cause of cancer death in Africa and the second cause of most common cancers in Cameroon. Herein, we determined the prevalence of high-risk HPV types in women and associated cervical cytologic abnormalities in Yaounde, Cameroon. Methodology: A cross-sectional study targeting HPV-positive women aged 20 and over was conducted between March and June 2020 at the Saint Martin de Porres’ Health Centre in Yaounde. HPV tests were performed by PCR for detection of HPVs 16, 18, 33, and 45. The test was performed on 616 women using exfoliated cell specimens; then, we processed on cytological diagnosis with Pap smears on HPV positive specimens. Results: The HPV types tested were detected in 137 participants, of which 38.7% with multiple HPV infections, and the remaining part with single HPV infections of type HPV 16 (28.5%), HPV 18 (17.5%), HPV 33 (10.2%), and HPV 45 (5.1%). Cervical cytologic abnormalities were found in 69.34% of participants including: LSIL (49.63%), HSIL (15.32%), ASC-US (3.66%) and AGC (0.73%). Co-infections with HPV 16 and HPV 18 were significantly associated with HSIL (p = 0.001) lesions, while HPV 45 was more common in participants with normal cytology (p = 0.001). Cervical lesion occurrence was significantly associated with the number of sexual partners (p = 0.02) and history of oral contraceptive pill use (p = 0.001). Conclusions: Our results suggest that HPV 16 and 18 are predominant in Yaounde, and are associated with more severe precancerous lesions.


2016 ◽  
Vol 14 (5) ◽  
pp. 808-815 ◽  
Author(s):  
Ferdaous Maâtouk ◽  
Mouna Maâtouk ◽  
Karima Bekir ◽  
Houcine Barhoumi ◽  
Abderrazak Maaref ◽  
...  

In this work we report the development of an electrochemical DNA biosensor with high sensitivity for mercury ion detection. A new matrix based on gold nanoparticles (AuNPs)-glutathione (GSH)/cysteine was investigated. The interaction between DNA oligonucleotides and Hg2+ ions followed by the formation of Thymine–Hg2+–Thymine (T–Hg2+–T) structures was quantified using different electrochemical methods. It has been shown that the electrochemical impedance spectroscopy (EIS) measurements and the differential pulse voltammetry (DPV) confirmed the specific interaction between the oligonucleotide receptor layer and the Hg2+ ions. Besides, the developed sensor exhibited high sensitivity towards mercury among some examined metal ions such as Pb2+, Cu2+ and Cd2+. As a result, a high electrochemical response and low detection limit of 50 pM were estimated in the case of Hg2+ ions. The developed DNA biosensor was applied successfully to the determination of Hg2+ions in wastewater samples.


Author(s):  
Daniel Duarte ◽  
Rodrigo Vieira ◽  
Elza Brito ◽  
Maria Pinheiro ◽  
Jeniffer Monteiro ◽  
...  

Purpose The aim of this study was to evaluate the overall and type-specific prevalence of human papillomavirus (HPV) infection among females living in riverside communities in the state of Pará, in the Eastern Brazilian Amazon. These communities are inhabited by low-income people, and are accessible only by small boats. Cervical cytology and risk factors for HPV infection were also assessed. Methods Cervical samples from 353 women of selected communities were collected both for Papanicolau (Pap) test and HPV detection. Conventional polymerase chain reaction (PCR) and real-time PCR were used to assess the overall and type-specific prevalence of HPV-16 and HPV-18, the main oncogenic types worldwide. Epidemiological questionnaires were used for the assessment of the risk factors for HPV infection. Results The mean age of the participants was 37 years (standard deviation [SD] ± 13.7). Most were married or with a fixed sexual partner (79%), and had a low educational level (80%) and family monthly income (< U$ 250; 53%). Overall, HPV prevalence was 16.4% (n = 58), with 8 cases of HPV-16 (2.3%) and 5 of HPV-18 (1.4%). Almost 70% of the women surveyed had never undergone the Pap test. Abnormal cytology results were found in 27.5% (n = 97) of the samples, with higher rates of HPV infection according to the severity of the lesions (p = 0.026). Conclusions The infections by HPV-16 and HPV-18 were not predominant in our study, despite the high prevalence of overall HPV infection. Nevertheless, the oncogenic potential of these types and the low coverage of the Pap test among women from riverside communities demonstrate a potential risk for the development of cervical lesions and their progression to cervical cancer, since the access to these communities is difficult and, in most cases, these women do not have access to primary care and public health services.


RSC Advances ◽  
2016 ◽  
Vol 6 (80) ◽  
pp. 76214-76222 ◽  
Author(s):  
Manvi Tak ◽  
Vinay Gupta ◽  
Monika Tomar

The article focuses on the synthesis of ZnO and CNTs based electrochemical DNA biosensor and its application towards meningitis DNA detection with high sensitivity as well as selectivity.


2005 ◽  
Vol 86 (12) ◽  
pp. 3235-3241 ◽  
Author(s):  
Marc Fiedler ◽  
Sigrun Ressler ◽  
Beatriz Campo-Fernández ◽  
Andreas Laich ◽  
Lars Jansen ◽  
...  

E7 proteins are major oncoproteins of high-risk human papillomaviruses (HPVs), which play a key role in cervical carcinogenesis. These proteins have been shown to immortalize primary human cells. Due to the absence of antibodies with suitable sensitivity and specificity, little is known about expression of the E7 oncoproteins in naturally infected tissues. Recently, high-level expression of the E7 protein of HPV-16, the most prevalent oncogenic HPV type, was demonstrated in cervical carcinomas by immunohistochemistry; however, approximately 15 additional high-risk HPV types are known to be associated with cervical carcinoma. It is unknown whether the E7 oncoproteins of HPV-18 and -45, the second and third most prevalent HPV types, are expressed in cervical cancers. Using antibodies against HPV-18 and -45 E7 proteins, it is shown here for the first time that the HPV-18 and -45 E7 proteins can be detected in cervical carcinoma biopsies. Together with anti-HPV-16 E7 antibodies, this could create the possibility of detecting E7 oncoproteins in approximately 80 % of all cervical cancers.


Author(s):  
Suzanna P Mongan ◽  
Andrijono Andrijono ◽  
Hartono Tjahadi

Objective: To determine whether HPV L1 gene methylation can be used in triage of precancerous cervical lesions. The main objective is to determine the genotype of HPV in cervical precancerous lesions and to determine the percentage, the sensitivity, specificity, positive predictive value, negative predictive value, and likelihood ratio of DNA HPV L1 methylation in precancerous cervical lesions. Methods: A number of 57 samples of paraffin blocks (FFPE) from precancerous lesions and cervical cancer biopsies in the Department of Pathology Faculty of Medicine-Cipto Mangunkusumo General Hospital that had been re-evaluated by the pathologist, underwent extraction of HPV DNA. The genotypes of HPV DNA were examined using primers GP5 / 6 and specific HPV 16, HPV 18 and HPV 52 probes and analyzed by real time PCR. Sequencing was performed on samples with unknown HPV DNA type that were detected using the specific probes to determine the type of HPV. Bisulfite conversion procedure was then performed for the samples that met the inclusion criteria. Results: There were 30 samples (52.6%) with CIN 1, 12 samples (21.1%) CIN 2, 9 samples (15.8%) CIN 3 and 6 samples (10.5%) of cervical cancer. Most of the samples were 36-45 years (35.1%). Of the total 57 samples, 55 samples were successfully extracted and determined the DNA genotyping of HPV (96.5%). HPV 16 infections both in the form of single or multiple was found to be 76.36%. The samples were mostly dominated by co-infection of HPV16 and 18 (49.1%) followed by HPV 16 (24.6%) and HPV 18 (14.0%). Based on the sequencing results there were other types of high risk HPV infection found: HPV 33, HPV 35, HPV 58 and also undeterminate risk HPV 53 and low risk HPV 54. After several procedures of optimization for methylation examination of HPV DNA L1 there was thin band found in electrophoresis procedure in 8 of 42 samples (19%) of HPV 16 after bisulfite conversion but once it was purified there weren’t any band found so we can not proceed to the stage sequencing. Until now we are still in the stage of optimizing the methylation procedure. Conclusion: HPV 16 infection were most commonly found in the form of single or multiple. Co-infection of HPV 16 and 18 were found in the majority of the samples. There were no significant correlation between HPV type and the severity of cervical lesions. Until now, the examination of DNA methylation HPV L1 already obtained eight samples of HPV 16 with a thin band on electrophoresis but the result could not be concluded because it is still in the process of optimization. [Indones J Obstet Gynecol 2017; 5-2: 120-126] Keywords: HPV DNA genotype, L1 gene methylation, precancerous cervical lesions


2020 ◽  
Author(s):  
Rodrigo Lopes da Silva ◽  
Zulmira da Silva Batista ◽  
Gerusinete Rodrigues Bastos ◽  
Ana Paula Almeida Cunha ◽  
Fábio Vidal Figueiredo ◽  
...  

Abstract Background: Cervical cancer is the fourth most common type of cancer affecting women globally. In Brazil, it is the third most frequent type of cancer in women and HPV is present in approximately 90% of cases. Evidence suggests that variants of HPV 16 can interfere biologically and etiologically during the development of cervical cancer. Methods: Cervix tumor fragments were collected, their DNA was extracted, and nested PCR was used to detect HPV. Positive samples were sequenced to determine the viral genotype. To characterize the HPV 16 strains, positive samples PCR was used to amplify the LCR and E6 regions of the HPV 16 virus. Results: Data from 120 patients with cervical cancer were analyzed. Most women were between 41 and 54 years of age, had schooling until primary school, a family income between 1 and 2 times the minimum wage and were married/in a consensual union. There was no statistically significant association between HPV or socio-demographic variables and risk factors for cervical cancer (P < 0.05). HPV was present in 88 women (73%). The most prevalent types were HPV 16 (47/54%), HPV 18 (12/13.8%), HPV 35 (6/6.9%) and HPV 45 (5/5.7%). Of the 47 HPV 16 positive cases, variant A (49%) was present in 23 samples, followed by variant D in 20 cases (43%), and variants B and C in 2 cases each (4%).The most prevalent histological type of HPV 16 tumors was epidermoid carcinoma, followed by adenocarcinoma. There was a statistically significant association between HPV 16 variants and the tumors’ histological types (P < 0.001).Conclusions: Knowledge of HPV 16 variants will provide data on their influence on the pathological and oncogenic aspects of cervical lesions.


2020 ◽  
Author(s):  
Rodrigo Lopes da Silva ◽  
Zulmira da Silva Batista ◽  
Gerusinete Rodrigues Bastos ◽  
Ana Paula Almeida Cunha ◽  
Fábio Vidal Figueiredo ◽  
...  

Abstract Background: Cervical cancer is the fourth most common type of cancer affecting women globally. In Brazil, it is the third most frequent type of cancer in women and HPV is present in approximately 90% of cases. Evidence suggests that variants of HPV 16 can interfere biologically and etiologically during the development of cervical cancer. Methods: Cervix tumor fragments were collected, their DNA was extracted, and nested PCR was used to detect HPV. Positive samples were sequenced to determine the viral genotype. To characterize the HPV 16 strains, positive samples PCR was used to amplify the LCR and E6 regions of the HPV 16 virus.Results: Data from 120 patients with cervical cancer were analyzed. Most women were between 41 and 54 years of age, had schooling until primary school, a family income between 1 and 2 times the minimum wage and were married/in a consensual union. There was no statistically significant association between HPV or socio-demographic variables and risk factors for cervical cancer (P < 0.05). HPV was present in 88 women (73%). The most prevalent types were HPV 16 (47/54%), HPV 18 (12/13.8%), HPV 35 (6/6.9%) and HPV 45 (5/5.7%). Of the 47 HPV 16 positive cases, variant A (49%) was present in 23 samples, followed by variant D in 20 cases (43%), and variants B and C in 2 cases each (4%).The most prevalent histological type of HPV 16 tumors was epidermoid carcinoma, followed by adenocarcinoma. There was a statistically significant association between HPV 16 variants and the tumors’ histological types (P < 0.001).Conclusions: Knowledge of HPV 16 variants will provide data on their influence on the pathological and oncogenic aspects of cervical lesions.


2006 ◽  
Vol 16 (3) ◽  
pp. 1041-1047
Author(s):  
P. Piña-Sánchez ◽  
D. M. Hernández-Hernández ◽  
R. LÓPEZ-ROMERO ◽  
G. VÁZQUEZ-ORTÍZ ◽  
C. PÉREZ-PLASENCIA ◽  
...  

Cervical cancer (CC) is the most common in Mexican female population. The human papillomavirus (HPV) 16 and 18 frequencies in worldwide may be different due to geographical distribution. We analyzed the prevalence of HPV types and determinated their association in cervical lesion in a Mexican population. One hundred fifty-nine normal cervical smears, 95 low-grade squamous intraepithelial lesions (LGSIL), 59 high-grade squamous intraepithelial lesions (HGSIL), and 108 CC samples of the patients were collected. HPV types were determined by sequencing. We detected 11 high-risk types, four low-risk types, three not determinated, and two probably high risk. HPV were present in 12%, 57%, 88%, and 92% from normal, LGSIL, HGSIL, and CC samples, respectively. HPV 16 was the most common in all cervical lesions (71.6% in CC). HPV 58 was present in 18.6% of HGSIL, and the HPV 18 in 4.6% of CC. The 76% of all detected viruses belong to A9 species branch. Control women showed high percentage of HPV high-risk infection, suggesting that this is a high-risk group. High frequency of HPV 16 compared with a low incidence of HPV 18 was observed. HPV 58 is frequently detected in HGSIL but low frequency is found in CC. These findings might be considered for HPV screening.


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