scholarly journals Rapid and ultrasensitive detection of circulating human papillomavirus E7 cell-free DNA as a cervical cancer biomarker

2020 ◽  
pp. 153537022097889
Author(s):  
Phetploy Rungkamoltip ◽  
Sasithon Temisak ◽  
Kitiya Piboonprai ◽  
Deanpen Japrung ◽  
Pattanapong Thangsunan ◽  
...  
Keyword(s):  
Cervical Cancer ◽  
Human Papillomavirus ◽  
Invasive Cervical Cancer ◽  
Digital Pcr ◽  
Clinical Samples ◽  
Lateral Flow Strip ◽  
Cell Free Dna ◽  
Hpv Dna ◽  
Hpv E7 ◽  
Free Dna

Circulating cell-free DNA (cfDNA) has attracted attention as a non-invasive biomarker for diagnosing and monitoring various cancers. Given that human papillomavirus (HPV) DNA integration and overexpression of E6/E7 oncogenes are pivotal events for carcinogenesis, we sought to determine if HPV E7 cfDNA could serve as a specific biomarker for cervical cancer detection. We applied droplet digital PCR (ddPCR) to quantify HPV16/18 E7 cfDNA from the serum of patients with cervical cancer, cervical intraepithelial neoplasia, and controls. HPV16/18 E7 cfDNA was highly specific for cervical cancer, displaying 30.77% sensitivity, 100% specificity, and an area under the curve of 0.65. Furthermore, we developed a sensitive isothermal detection of HPV16/18 E7 and the PIK3CA WT reference gene based on recombinase polymerase amplification combined with a lateral flow strip (RPA-LF). The assay took less than 30 min and the detection limit was 5–10 copies. RPA-LF exhibited 100% sensitivity and 88.24% specificity towards HPV16/18 E7 cfDNA in clinical samples. The agreement between RPA-LF and ddPCR was 83.33% ( κ = 0.67) for HPV16 E7 and 100% ( κ = 1.0) for HPV18 E7, indicating a good correlation between both tests. Therefore, we conclude that HPV E7 cfDNA represents a potential tumor marker with excellent specificity and moderate sensitivity for minimally invasive cervical cancer monitoring. Moreover, the RPA-LF assay provides an affordable, rapid, and ultrasensitive tool for detecting HPV cfDNA in resource-limited settings.

Evaluation of Cervical Cancer Screening Tools; INNO-LiPA® HPV Genotyping Extra-II Assay versus E7/E6 oncoproteins, How is reliable and practical?

2021 ◽  
Vol 15 (5) ◽  
pp. 1276-1281
Author(s):  
R. Bahramabadi ◽  
M. K. Arababadi ◽  
M. Iranpour ◽  
E. Mohebbi ◽  
Z. Honarvar ◽  
...  
Keyword(s):  
Cervical Cancer ◽  
Human Papillomavirus ◽  
Western Blot ◽  
Pap Smear ◽  
Diagnostic Methods ◽  
Screening Tools ◽  
Hpv Dna ◽  
Hpv Genotyping ◽  
Hpv E7 ◽  
Diagnostic Values

Background: High-Risk Human papillomavirus (HR-HPV) has been well established as the cervical cancer (CC) risk factor. In recent years, various diagnostic methods of human papillomaviruses (HPV) have been developed to promote sensitivity and specificity of CC screening which leads to a low mortality rate. This study aimed to compare diagnostic test metrics of two HPV diagnostic techniques, including Western blot and INNO-LiPA HPV Genotyping Extra II assay methods in asymptomatic or subclinical patients, among the South-Eastern Iranian women. Methods: 323 women were referred to the Pathology and Stem Cell Research Center, from February 2018 to January 2020. HPV-DNA with the INNO-LiPA HPV Genotyping Extra-II Assay kit and the western blot assays for HPV E7 and E6 assessment were employed. Results: Overall, 163 (50.4%) samples were dysplastic pap smear, the specificity of the HPV DNA test by INNO-LiPA HPV Genotyping Extra-II Assay test was significantly higher than the E7/E6 oncoproteins finding (67.3 vs. 49.9%), and the sensitivity was lower (96.6 vs. 74.8%), respectively. Conclusions: HR-HPV E7/E6 oncoproteins expression was evaluated as a possible novel biomarker for CC screening in pap smear as the preliminary test with satisfactory diagnostic values for HR-HPV types 16 and 18. The corresponding diagnostic values may be further improved by combining HPV DNA tests with the INNO-LiPA HPV Genotyping Extra-II test. Also, they may prove helpful for HR-HPV infection diagnosis in cases that the patients are asymptomatic or subclinical. Keywords: Cervical Cancer; Human Papillomavirus (HPV); Diagnostic Screening Programs; Oncogene Proteins

Human papillomavirus, coinfection with Schistosoma hematobium, and cervical neoplasia in rural Tanzania

2003 ◽  
Vol 13 (4) ◽  
pp. 505-509 ◽  
Author(s):  
K. U. Petry ◽  
U. Scholz ◽  
B. Hollwitz ◽  
R. Von Wasielewski ◽  
C. J.L.M. Meijer
Keyword(s):  
Cervical Cancer ◽  
Human Papillomavirus ◽  
Invasive Cervical Cancer ◽  
Clinical History ◽  
Hpv Infection ◽  
Separate Analysis ◽  
Hpv Dna ◽  
Rural Tanzania ◽  
Increased Risk ◽  
Local Immunosuppression

Cervical cancer is the most common malignant tumor among women in Tanzania and other countries in tropical Africa. Genital schistosomiasis has been proposed as a possible cofactor in the genesis of this malignant disease that might contribute to its high incidence in regions where bilharzias is endemic. One hundred nine Tanzanian patients from an area with endemic bilharzias who were transferred to a gynecologic out-patient clinic were age-matched with 109 German controls. In patients and controls, separate samples were taken for cytologic assessment and human papillomavirus (HPV) DNA detection using the Hybrid Capture 2 assay (HC2) and PCR (GP5+/6 +). Samples that tested positive for HPV DNA with general primers were re-tested with HPV type-specific primers. After application of 3% acetic acid, punch biopsies were taken from any cervical lesion. Patients were interviewed for recent symptoms or clinical history suggestive of bilharzias. Urine samples from all patients were examined for the presence of schistosoma hematobium ova. Additionally six Tanzanian patients with invasive cervical cancer were included for separate analysis. Patients and controls had an identical prevalence of HPV-DNA (21.5%) using HC2. Based on PCR results with general primers, the corresponding prevalence was 34.5% for Tanzanian cases and 26.9% for German controls. A history suggestive of bilharzias and/or active schistosomiasis were associated with a significantly increased risk for infection with high-risk HPV types. We conclude that infection with Schistosoma hematobium seems to favor persistent genital HPV infection either by traumatizing the genital epithelium and/or by local immunosuppression.

scholarly journals Droplet Digital PCR Diagnosis of Human Schistosomiasis: Parasite Cell-Free DNA Detection in Diverse Clinical Samples

2017 ◽  
Vol 216 (12) ◽  
pp. 1611-1622 ◽  
Author(s):  
Kosala G Weerakoon ◽  
Catherine A Gordon ◽  
Gail M Williams ◽  
Pengfei Cai ◽  
Geoffrey N Gobert ◽  
...  
Keyword(s):  
Dna Detection ◽  
Digital Pcr ◽  
Droplet Digital Pcr ◽  
Clinical Samples ◽  
Cell Free Dna ◽  
Parasite Cell ◽  
Free Dna ◽  
Human Schistosomiasis ◽  
Pcr Diagnosis

scholarly journals Evaluation of pre-analytical factors affecting plasma DNA analysis

2017 ◽  
Author(s):  
Havell Markus ◽  
Tania Contente-Cuomo ◽  
Winnie S. Liang ◽  
Mitesh J. Borad ◽  
Shivan Sivakumar ◽  
...  
Keyword(s):  
Dna Analysis ◽  
Fragment Size ◽  
Digital Pcr ◽  
Clinical Samples ◽  
Blood Collection ◽  
Plasma Samples ◽  
Plasma Dna ◽  
Cell Free Dna ◽  
Free Dna ◽  
The Impact

AbstractPre-analytical factors can significantly affect circulating cell-free DNA (cfDNA) analysis. However, there are few robust methods to rapidly assess sample quality and the impact of pre-analytical processing. To address this gap and to evaluate effects of DNA extraction methods and blood collection tubes on cfDNA yield and fragment size, we developed a multiplexed droplet digital PCR (ddPCR) assay with 5 short and 4 long amplicons targeting single copy genomic loci (mean amplicon size: 71 bp and 471 bp respectively). Using this assay, we compared performance of 7 cfDNA extraction kits and found cfDNA yield and fragment size varies significantly between them. We also compared 3 blood collection protocols used to collect plasma samples from 23 healthy volunteers (EDTA tubes processed within 1 hour and Cell-free DNA BCT tubes at ambient temperature processed within 24 hours and 72 hours of collection). To assess whether cell-stabilizing preservative in BCT tubes introduced noise in cfDNA, we performed digital targeted sequencing. We found no significant differences in cfDNA yield, fragment size and background sequencing noise between these protocols. In 219 clinical samples tested for quality using the ddPCR assay, cfDNA fragment size was significantly shorter in plasma samples immediately processed for ctDNA analysis compared to archived samples, suggesting background DNA contributed by lysed peripheral blood cells. In summary, we describe a multiplexed ddPCR approach that enables cfDNA quality assessment and could inform the design of future circulating tumor DNA studies.Gene namesNone

scholarly journals Human Hsp70 and Hsp40 Chaperone Proteins Facilitate Human Papillomavirus-11 E1 Protein Binding to the Origin and Stimulate Cell-free DNA Replication

1998 ◽  
Vol 273 (46) ◽  
pp. 30704-30712 ◽  
Author(s):  
Jen-Sing Liu ◽  
Shu-Ru Kuo ◽  
Alexander M. Makhov ◽  
Douglas M. Cyr ◽  
Jack D. Griffith ◽  
...  
Keyword(s):  
Human Papillomavirus ◽  
Dna Replication ◽  
Protein Binding ◽  
Chaperone Proteins ◽  
Cell Free Dna ◽  
E1 Protein ◽  
Free Dna ◽  
Stimulate Cell ◽  
Human Hsp70

Human papillomavirus (HPV) genotyping by HPV DNA chip in cervical cancer and precancerous lesions

2005 ◽  
Vol 15 (1) ◽  
pp. 81-87 ◽  
Author(s):  
G.-Y. Lee ◽  
S.-M. Kim ◽  
S.-Y. Rim ◽  
H.-S. Choi ◽  
C.-S. Park ◽  
...  
Keyword(s):  
Cervical Cancer ◽  
Human Papillomavirus ◽  
Precancerous Lesions ◽  
Dna Chip ◽  
Hpv Dna ◽  
Hpv Genotyping

scholarly journals Human papillomavirus type 16 genomic variation in women with subsequent in situ or invasive cervical cancer: prospective population-based study

2018 ◽  
Vol 119 (9) ◽  
pp. 1163-1168 ◽  
Author(s):  
Laila Sara Arroyo-Mühr ◽  
Camilla Lagheden ◽  
Emilie Hultin ◽  
Carina Eklund ◽  
Hans-Olov Adami ◽  
...  
Keyword(s):  
Cervical Cancer ◽  
Human Papillomavirus ◽  
Invasive Cervical Cancer ◽  
Population Based ◽  
Genomic Variation ◽  
Population Based Study ◽  
Human Papillomavirus Type 16

scholarly journals Antibodies against Early Proteins of Human Papillomaviruses as Diagnostic Markers for Invasive Cervical Cancer

1998 ◽  
Vol 36 (2) ◽  
pp. 475-480 ◽  
Author(s):  
Wolfgang Meschede ◽  
Klaus Zumbach ◽  
Joris Braspenning ◽  
Martin Scheffner ◽  
Luis Benitez-Bribiesca ◽  
...  
Keyword(s):  
Cervical Cancer ◽  
Invasive Cervical Cancer ◽  
Human Papillomaviruses ◽  
Antibody Detection ◽  
Diagnostic Tools ◽  
Immunological Monitoring ◽  
Hpv Dna ◽  
Native Proteins ◽  
Human Sera ◽  
E6 And E7

Cervical cancer is the most prevalent tumor in developing countries and the second most frequent cancer among females worldwide. Specific human papillomaviruses (HPVs) and, most notably, HPV types 16 and 18 are recognized as being causally associated with this malignancy. Antibodies against early HPV proteins E6 and E7 have been found more often in patients with tumors than in controls. Existing peptide enzyme-linked immunosorbent assays (ELISAs) for the detection of anti-E6 and anti-E7 antibodies in human sera have low levels of sensitivity and specificity and thus are not suitable for use as diagnostic tools. Based on highly purified recombinant native proteins, we developed four sandwich ELISAs for the detection of antibodies against HPV type 16 and 18 E6 and E7 proteins. We demonstrate their sensitivities and high degrees of specificity for cervical cancer. Among a total of 501 serum specimens from unselected patients with invasive cervical cancer, 52.9% reacted positively in at least one of the four assays. In contrast, among 244 serum specimens from control subjects without cervical cancer, only 2 reactive serum specimens (0.8%) were found. For 19 of 19 antibody-positive patients, the HPV type indicated by seroreactivity was identical to the HPV DNA type found in the tumor, which also indicates a high degree of specificity for antibody detection with respect to HPV type. In a direct comparison of 72 serum specimens from patients with cervical cancer, 56% of the specimens reacted in at least one of the four protein ELISAs, whereas 40% reacted in at least one of seven peptide ELISAs covering the four antigens. These assays could be of value for the detection of invasive cervical cancer in settings in which cytology-based early tumor screening is not available, for the clinical management of patients diagnosed with cervical cancer, and for the immunological monitoring of E6 and E7 vaccination trials.

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