A New Sandwich ELISA Test Simultaneously Detecting E7 Proteins of HPV-16, 18 and 45 in Cervical Smears

Metzger, Christina; Pittl, Annabella; Kaufmann, Andreas M.; Αγοραστός, Θεόδωρος; Chatzistamatiou, Κimon; Böcher, Oliver; Zwerschke, Werner; Pircher, Haymo; Koch, Isabel; Jansen‐Dürr, Pidder

doi:10.4172/2327-5073.1000260

Cited by 5 publications

(4 citation statements)

References 25 publications

(31 reference statements)

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“…As known, E6 and E7 transforming proteins are consistently expressed in HPV life cycle [39–41]. Several studies have shown the induction of antibody responses against E6 and E7 oncoproteins in patients with cervical cancer higher than in healthy subjects [39, 42, 43]. These studies and other reports usually indicated HPV seroreactivity for cancer patients in early or late stage; but our study showed a high seroreactivity in women-exposed to high-risk HPV types (i.e.…”

Section: Discussionmentioning

confidence: 99%

Significance of serum antibodies against HPV E7, Hsp27, Hsp20 and Hp91 in Iranian HPV-exposed women

et al. 2019

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BackgroundAmong different types of human papillomavirus (HPV), types 16 and 18 were known to be high-risk agents causing mainly cervical cancer. Up to now, the potential of HPV E7 protein has been proved as a diagnostic marker of cervical cancer. Moreover, the levels of anti-heat shock protein (Hsp) and anti-high mobility group box-1 (HMGB1) antibodies in cancer patients have been useful in tumor diagnosis. The goal of the present study was to determine the efficiency of the potential serologic markers including HPV E7, Hsp20, Hsp27 proteins and Hp91 peptide in Iranian HPV-exposed women, for the first time.MethodsAt first, the recombinant HPV E7, Hsp20 and Hsp27 proteins were expressed in E. coli system, and purified by affinity chromatography under native conditions. Then, antibody responses were detected against the recombinant proteins as well as Hp91 peptide as potential markers in 49 Iranian women who were seropositive for HPV-16 and 18 L1 capsids (i.e., HPV-exposed women) and 49 controls using indirect ELISA.ResultsOur data indicated that the seroreactivities of women exposed to HPV16, HPV18 and both of them against the recombinant E7, Hsp20, Hsp27 proteins and Hp91 peptide were significantly higher than those in control group (p < 0.05 for HPV16 or HPV18; p < 0.01 for both of them versus all markers). HPV-exposed women with high antibody responses to HPV-16 and 18 L1 capsids as a commercial biomarker had significant seroreactivity to HPV-16 and 18 E7 and Hsp27 (p < 0.05). The recombinant E7 and Hsp27 proteins showed higher efficiency than Hsp20 and Hp91 for detection of individuals exposed to HPV infections (p < 0.05).ConclusionGenerally, the levels of serum E7 and Hsp27 were increased in HPV-16 and 18 L1- seropositive women suggesting their potential value as a diagnostic marker for HPV infections.Electronic supplementary materialThe online version of this article (10.1186/s12879-019-3780-2) contains supplementary material, which is available to authorized users.

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Section: Discussionmentioning

confidence: 99%

Significance of serum antibodies against HPV E7, Hsp27, Hsp20 and Hp91 in Iranian HPV-exposed women

et al. 2019

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“…Detection of hrHPVE7 protein was performed via a hrHPVE7 Sandwich ELISA which was developed in a collaboration between the laboratories of Innsbruck University and Mikrogen GmbH, Neuried, Germany during the PIPAVIR project (Fig. ).…”

Section: Methodsmentioning

confidence: 99%

Human papillomavirus E7 protein detection as a method of triage to colposcopy of HPV positive women, in comparison to genotyping and cytology. Final results of the PIPAVIR study

et al. 2017

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The objective of the presented cross-sectional-evaluation-screening study is the clinical evaluation of high-risk(hr)HPVE7-protein detection as a triage method to colposcopy for hrHPV-positive women, using a newly developed sandwich-ELISA-assay. Between 2013-2015, 2424 women, 30-60 years old, were recruited at the Hippokratio Hospital, Thessaloniki/Greece and the Im Mare Klinikum, Kiel/Germany, and provided a cervical sample used for Liquid Based Cytology, HPV DNA genotyping, and E7 detection using five different E7-assays: "recomWell HPV16/18/45KJhigh", "recomWell HPV16/18/45KJlow", "recomWell HPV39/51/56/59", "recomWell HPV16/31/33/35/52/58" and "recomWell HPVHRscreen" (for 16,18,31,33,35,39,45,51,52,56,58,59 E7), corresponding to different combinations of hrHPVE7-proteins. Among 1473 women with eligible samples, those positive for cytology (ASCUS+ 7.2%), and/or hrHPV DNA (19.1%) were referred for colposcopy. Cervical Intraepithelial Neoplasia grade 2 or worse (CIN2+) was detected in 27 women (1.8%). For HPV16/18-positive women with no triage, sensitivity, positive predictive value (PPV) and the number of colposcopies needed to detect one case of CIN2+ were 100.0%, 11.11% and 9.0 respectively. The respective values for E7-testing as a triage method to colposcopy ranged from 75.0-100.0%, 16.86-26.08% and 3.83-5.93. Sensitivity and PPV for cytology as triage for hrHPV(non16/18)-positive women were 45.45% and 27.77%; for E7 test the respective values ranged from 72.72-100.0% and 16.32-25.0%. Triage of HPV 16/18-positive women to colposcopy with the E7 test presents better performance than no triage, decreasing the number of colposcopies needed to detect one CIN2+. In addition, triage of hrHPV(non16/18)-positive women with E7 test presents better sensitivity and slightly worse PPV than cytology, a fact that advocates for a full molecular screening approach.

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“…The recom Well Keratin 5/8/18 ELISA has potential to serve as a test for sample validation in a range of cervical cancer screening tests. Future prospects include ELISAs developed for detection of elevated tumor suppressor protein p16, and HPV oncoprotein E7 [11,21,39,40]. A dual staining cytology technique for p16 and Ki-67 shows much promise [12].…”

Section: Discussionmentioning

confidence: 99%

Development of a novel enzyme-linked immunosorbent assay for the detection of cytokeratins 5, 8, and 18, and its implications in cervical cancer screening standardization

McNamara¹,

Kaufmann²,

Vasiljeva³

et al. 2018

J Clin Mol Med

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There is a dire need for improved cervical cancer screening methods. New tests are in the pipeline, but their diagnostic capabilities may be limited without a way to assess specimen validity. Here we describe an enzyme-linked immunosorbent assay (ELISA) that captures specific intermediate filament proteins (cytokeratins) from potentially transformable target cells located within or originating from the cervical transformation zone as a means of cervical specimens. Seventy-five uterine cervical samples negative for human papillomavirus (HPV) were grouped based on microscopic analysis for the presence or absence of cervical transformation zone cells, and all samples were tested in both a newly developed recom Well Keratin 5/8/18 ELISA and a pan keratin control ELISA. Additionally, 15 samples from HPVpositive patients manifesting histopathologic lesions or squamous cell carcinoma were tested. Our results demonstrate the presence and detectability by ELISA of keratins 5, 8, and 18 in parabasal, squamous metaplastic, and endocervical cells, while simultaneously suggesting their absence in differentiated squamous cells. We also validate with ELISA the expression of these keratins in HPV-induced disease-state individuals. Our findings indicate recom Well Keratin 5/8/18 ELISA may be useful as a standardizing tool in cervical cancer screening, or alternatively as a quality indicator to denote specimen adequacy.

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A New Sandwich ELISA Test Simultaneously Detecting E7 Proteins of HPV-16, 18 and 45 in Cervical Smears

Cited by 5 publications

References 25 publications

Significance of serum antibodies against HPV E7, Hsp27, Hsp20 and Hp91 in Iranian HPV-exposed women

Significance of serum antibodies against HPV E7, Hsp27, Hsp20 and Hp91 in Iranian HPV-exposed women

Human papillomavirus E7 protein detection as a method of triage to colposcopy of HPV positive women, in comparison to genotyping and cytology. Final results of the PIPAVIR study

Development of a novel enzyme-linked immunosorbent assay for the detection of cytokeratins 5, 8, and 18, and its implications in cervical cancer screening standardization

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