scholarly journals Impact of specimen age on its DNA quality for Formalin-Fixed Paraffin-Embedded HPV specimens

2018 ◽  
Author(s):  
James Yeongjun Park
Keyword(s):  
Molecular Profiling ◽  
Molecular Characteristics ◽  
Dna Quality ◽  
Formalin Fixed Paraffin ◽  
Formalin Fixed Paraffin Embedded ◽  
Hpv Types ◽  
The Relationship ◽  
Long Term Preservation ◽  
Formalin Fixed

AbstractFormalin fixation and paraffin embedding (FFPE) allows the storage of diagnostic and surplus tissue in archival banks. Therefore, FFPE is now a standard method for long-term preservation of tissue biopsies as FFPE of samples preserves the morphology of tissue. Unfortunately, the FFPE process engenders chemical changes and degradation in tissue macromolecules that can pose threats to reliable subsequent analysis. DNA, while more resistant to FFPE in comparison to RNA and protein, is also subject to such chemical formations and degradation. This study provides robust findings about the relationship between DNA quality and specimen age from 10252 FFPE HPV specimens. This paper suggests that there is a perceptible degrading effect in DNA quality as specimens age. This study suggests that the biospecimen may begin to take on new characteristics after certain storage years, and such changes may result in inaccurate determinations of the molecular characteristics of the biospecimen during analysis. Results from this study demonstrate that older HPV specimens are more inclined to be tested negative or inadequate from commercial HPV genotyping assays. Same findings are conferred when multiple genotyping assays are involved with HPV testing. Also, older HPV samples typify fewer HPV types compared to younger HPV samples. The results from this study will be useful to enhance potential scope for next fixation methods such as ethanol fixation that may be equally useful for both molecular profiling and histology as FFPE.

scholarly journals Quantitative radioimmunohistochemical measurements of p185(erbB-2) in frozen tissue sections.

1996 ◽  
Vol 44 (11) ◽  
pp. 1251-1259 ◽  
Author(s):  
J R Reeves ◽  
J J Going ◽  
G Smith ◽  
T G Cooke ◽  
B W Ozanne ◽  
...  
Keyword(s):  
Tumor Biology ◽  
Breast Carcinomas ◽  
Tissue Sections ◽  
Biopsy Specimens ◽  
Formalin Fixed Paraffin ◽  
Formalin Fixed Paraffin Embedded ◽  
Interstudy Variability ◽  
High Degree ◽  
The Relationship ◽  
Formalin Fixed

The relationship between expression of the c-erbB-2 proto-oncogene and the biology of breast cancer has been investigated widely, most studies using immunohistochemistry in formalin-fixed, paraffin-embedded tissues. This technique is at best semiquantitative and there is a high degree of interstudy variability because of its subjective nature and poor methodological standardization. The relationship between the levels of expression and biology can be examined thoroughly only with an accurately quantitative technique. We have developed a radioimmunohistochemical assay to measure p185(erbB-2) in tissue biopsy specimens. The method involves incubating frozen sections with 125I-labeled monoclonal antibody, microautoradiograpy, and grain counting with image analysis. Sections of cell pellets with known c-erbB-2 levels are processed with each batch of samples as internal calibration standards. We have quantified c-erbB-2 expression in 60 breast carcinomas and compared the results with conventional immunohistochemistry. Radioimmunohistochemistry measured receptor levels throughout the range of expression in breast carcinomas, whereas conventional immunohistochemistry detected the protein only in the highest expressing tumors. The quantitative, objective data produced by radioimmunohistochemistry allow a more thorough evaluation of the relationship between c-erbB-2 expression and tumor biology. This technique may have applications in other fields where quantitative data is required and relevant monoclonal antibodies are available.

scholarly journals Improved RT-PCR Amplification for Molecular Analyses with Long-term Preserved Formalin-fixed, Paraffin-embedded Tissue Specimens

2006 ◽  
Vol 54 (7) ◽  
pp. 773-780 ◽  
Author(s):  
Kiyohiro Hamatani ◽  
Hidetaka Eguchi ◽  
Keiko Takahashi ◽  
Kazuaki Koyama ◽  
Mayumi Mukai ◽  
...  
Keyword(s):  
Pcr Amplification ◽  
Rt Pcr ◽  
Molecular Analyses ◽  
Formalin Fixed Paraffin ◽  
Formalin Fixed Paraffin Embedded ◽  
Tissue Specimens ◽  
Formalin Fixed

Immunohistochemical Detection of Canine Adenovirus in Paraffin Sections of Liver

1986 ◽  
Vol 23 (4) ◽  
pp. 478-484 ◽  
Author(s):  
P. M. Rakich ◽  
K. W. Prasse ◽  
P. D. Lukert ◽  
L. M. Cornelius
Keyword(s):  
Immunohistochemical Detection ◽  
Hepatic Inflammation ◽  
Paraffin Sections ◽  
Inflammatory Lesions ◽  
Long Term Stability ◽  
Formalin Fixed Paraffin ◽  
Wide Range ◽  
Formalin Fixed Paraffin Embedded ◽  
Formalin Fixed

An avidin-biotin immunoperoxidase procedure was optimized for detection of canine adenoviral antigens in formalin-fixed, paraffin-embedded liver. Long-term stability of viral antigen was shown by successful demonstration of virus in liver tissue preserved up to six years from dogs with infectious canine hepatitis. This immunohistochemical stain was applied to sections from livers with a wide range of inflammatory lesions. Examination of sections from 53 dogs yielded five livers with small amounts of adenovirus. An additional virus-positive liver was identified from a dog with no hepatic inflammation. Although a cause and effect relationship remains to be determined, these findings suggest a possible connection between canine adenovirus and spontaneous chronic hepatitis.

Evaluating the Feasibility of DNA Methylation Analyses Using Long-Term Archived Brain Formalin-Fixed Paraffin-Embedded Samples

2016 ◽  
Vol 55 (1) ◽  
pp. 668-681 ◽  
Author(s):  
Stine T. Bak ◽  
Nicklas H. Staunstrup ◽  
Anna Starnawska ◽  
Tina F. Daugaard ◽  
Jens R. Nyengaard ◽  
...  
Keyword(s):  
Dna Methylation ◽  
Formalin Fixed Paraffin ◽  
Formalin Fixed Paraffin Embedded ◽  
Formalin Fixed

Population-based p16 and HPV positivity rates in oropharyngeal cancer in Southeast Scotland

2015 ◽  
Vol 68 (10) ◽  
pp. 849-852 ◽  
Author(s):  
L A R Wells ◽  
E J Junor ◽  
B Conn ◽  
S Pattle ◽  
K Cuschieri
Keyword(s):  
Hpv Infection ◽  
Population Based ◽  
Papilloma Virus ◽  
P16 Immunohistochemistry ◽  
Formalin Fixed Paraffin ◽  
Formalin Fixed Paraffin Embedded ◽  
Hpv Types ◽  
Positive Groups ◽  
Formalin Fixed ◽  
Cause Specific Survival

We assessed a population-based cohort of patients diagnosed with oropharyngeal squamous cell carcinoma in Southeast Scotland over 13 months. p16 and human papilloma virus (HPV) expression were determined, and correlated with stage, treatment, smoking and alcohol history, and disease outcomes. Retrospective analysis was performed on 60 patients. p16 immunohistochemistry and HPV genotyping were performed on formalin-fixed paraffin-embedded tissues. HPV infection (as defined by p16 positivity and/or HPV PCR positivity) was identified in 57% of samples, while dual positives were detected in 45% of cases. HPV16 was most prevalent of the HPV types and was associated with 90% of positive samples. Cause-specific 1-year and 2-year survivals were 82.5% and 78.2%, respectively. The p16-positive and HPV-positive groups demonstrated significantly increased cause-specific survival in comparison with their negative counterparts.

Optimization of RNA extraction protocol for long-term archived formalin-fixed paraffin-embedded tissues of horses

2019 ◽  
Vol 110 ◽  
pp. 104289 ◽  
Author(s):  
Gisele Silva Boos ◽  
Daniel Nobach ◽  
Klaus Failing ◽  
Markus Eickmann ◽  
Christiane Herden
Keyword(s):  
Rna Extraction ◽  
Extraction Protocol ◽  
Formalin Fixed Paraffin ◽  
Formalin Fixed Paraffin Embedded ◽  
Formalin Fixed

Factors affecting immunoreactivity in long-term storage of formalin-fixed paraffin-embedded tissue sections

2015 ◽  
Vol 144 (1) ◽  
pp. 93-99 ◽  
Author(s):  
Federica Grillo ◽  
Simona Pigozzi ◽  
Paola Ceriolo ◽  
Paola Calamaro ◽  
Roberto Fiocca ◽  
...  
Keyword(s):  
Factors Affecting ◽  
Tissue Sections ◽  
Long Term Storage ◽  
Formalin Fixed Paraffin ◽  
Formalin Fixed Paraffin Embedded ◽  
Term Storage ◽  
Formalin Fixed

scholarly journals Quality assessment of long-term stored formalin-fixed paraffin embedded tissues for histopathological evaluation

2018 ◽  
Vol 31 (1) ◽  
pp. 61-64 ◽  
Author(s):  
Yuko Ono ◽  
Hiroko Sato ◽  
Toshiko Miyazaki ◽  
Katsuya Fujiki ◽  
Eisuke Kume ◽  
...  
Keyword(s):  
Quality Assessment ◽  
Formalin Fixed Paraffin ◽  
Histopathological Evaluation ◽  
Formalin Fixed Paraffin Embedded ◽  
Formalin Fixed

High-risk human papillomavirus (HPV) detection in formalin-fixed paraffin-embedded cervical tissues: performances of Aptima HPV assay and Beckton Dickinson (BD) Onclarity assay

2021 ◽  
pp. jclinpath-2021-207657
Author(s):  
Gozde Kir ◽  
Humeyra Gunel ◽  
Zeynep Cagla Olgun ◽  
W Glenn McCluggage
Keyword(s):  
Human Papillomavirus ◽  
High Risk ◽  
Hpv 16 ◽  
Formalin Fixed Paraffin ◽  
Formalin Fixed Paraffin Embedded ◽  
Ffpe Tissues ◽  
Hpv Types ◽  
High Risk Hpv ◽  
Hpv Assays ◽  
Formalin Fixed

AimThere are many scenarios where high-risk human papillomavirus (HPV) detection in formalin-fixed paraffin-embedded (FFPE) specimens is important. However, there is no Food and Drug Administration (FDA)-approved and clinically validated technique for detecting high-risk HPV in FFPE tissues. In this study, we evaluated two commercially available HPV assays which are FDA-approved for use on cytology specimens, the Aptima HPV assay and the Beckton Dickinson (BD) Onclarity assay, to detect high-risk HPV in FFPE tissues of cervical high-grade squamous intraepithelial lesion (HSIL) and squamous cell carcinoma (SCC).MethodsA total of 189 cases (46 SCC, 107 HSIL and 36 benign/normal) were tested for high-risk HPV with the Aptima HPV assay and a subset of cases (n=97) with the BD Onclarity assay.ResultsThe sensitivities of the Aptima and BD Onclarity HPV assays were 99.4% (95% CI 96.46% to 99.98%) and 75.9% (95% CI 65.27% to 84.62%), respectively; the specificity and positive predictive value (PPV) of the two assays were 100%. Negative predictive values of the Aptima and BD Onclarity HPV assays were 97.3% (95% CI 83.61% to 99.61%) and 67.7% (95% CI 58.91% to 75.47%), respectively. The kappa value (0.96) for comparison of the distribution of high-risk HPV types between the two assays was high. HPV 16 was the most common high-risk HPV type for HSIL and SCC cases. However, SCC cases had higher percentages of HPV 16 and HPV 18/45 and lower percentages of other high-risk HPV types compared with HSIL cases.ConclusionBoth assays are reliable methods for high-risk HPV detection and genotype determination in FFPE specimens, with high PPV and specificity. The Aptima HPV assay has the advantage of higher sensitivity. As far as we are aware, this is the first study comparing the Aptima HPV assay and the BD Onclarity assay in FFPE tissues. Our study results should be tested and confirmed in larger cohorts.

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