Risk for molecular contamination of tissue samples evaluated for targeted anti-cancer therapy
| العنوان: | Risk for molecular contamination of tissue samples evaluated for targeted anti-cancer therapy |
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| المؤلفون: | Ofer Ben-Izhak, Einav Simon, Dov Hershkovitz, Ibrahim Fahoum, Edmond Sabo, Eyal Asor, Michael Y. Stav |
| المصدر: | PLoS ONE, Vol 12, Iss 3, p e0173760 (2017) PLoS ONE |
| بيانات النشر: | Public Library of Science (PLoS), 2017. |
| سنة النشر: | 2017 |
| مصطلحات موضوعية: | 0301 basic medicine, Mutation rate, Lung Neoplasms, Tissue Fixation, Cancer Treatment, lcsh:Medicine, Artificial Gene Amplification and Extension, Pathology and Laboratory Medicine, medicine.disease_cause, Polymerase Chain Reaction, law.invention, Exon, 0302 clinical medicine, Mutation Rate, Limit of Detection, law, Medicine and Health Sciences, Molecular Targeted Therapy, EGFR Exon 21 Mutation, Pathology, Molecular, lcsh:Science, DNA extraction, Polymerase chain reaction, Mutation, Paraffin Embedding, Multidisciplinary, Exons, Laboratory Equipment, ErbB Receptors, Deletion Mutation, Oncology, 030220 oncology & carcinogenesis, Engineering and Technology, Molecular Pathology, Research Article, Equipment, Adenocarcinoma of Lung, Adenocarcinoma, Research and Analysis Methods, Deep sequencing, 03 medical and health sciences, Extraction techniques, Genetics, medicine, Point Mutation, Humans, Microtomes, Molecular Biology Techniques, Molecular Biology, business.industry, Point mutation, lcsh:R, Biology and Life Sciences, Molecular biology, 030104 developmental biology, lcsh:Q, business |
| الوصف: | With the increasing usage of sensitive PCR technology for pharmacogenetics, cross contamination becomes a significant concern. Researchers employed techniques which basically include replacing laboratory equipment after each sample preparation; however, there are no recommended guidelines. In the present work we wanted to evaluate the risk of cross contamination during tissue processing using the routine precaution measures. Twenty-one surgical samples of lung adenocarcinoma were used, of which 7 contained EGFR exon 19 mutation, 7 contained EGFR exon 21 mutation (p.L858R) and 7 were EGFR wild-type. The samples were ordered by alternating the mutation group to maximize the potential for cross contamination and underwent tissue sectioning and de-paraffinization. The entire process was performed using the same tools. Following DNA extraction all samples underwent PCR amplification and were scrutinized for small fractions of EGFR mutation using deep sequencing with the Ion torrent PGM technology. Twenty samples yielded results. The fraction of mutated copies was 41 ± 23% (range 11-66) for the cases with known exon 19 mutation and 48±24% (range 0-65) for the cases with known exon 21 mutations. No in-frame exon 19 deletion mutations were identified in the wild-type (WT) and exon 21 groups. The fraction of EGFR exon 21 (codon 858) mutations was 0.018±0.014% (range 0-0.05%) in the WT and exon 19 groups, which was not statistically different than the background sequencing artifact noise for the same base-pair alteration (p = 0.21). Our results suggest that standard precautions are sufficient for molecular pathology diagnosis of surgical samples and are not associated with increased risk of cross contamination. |
| اللغة: | English |
| تدمد: | 1932-6203 |
| الوصول الحر: | https://explore.openaire.eu/search/publication?articleId=doi_dedup___::a80ace011535ac71c55a861b195d97b2Test http://europepmc.org/articles/PMC5348008?pdf=renderTest |
| حقوق: | OPEN |
| رقم الانضمام: | edsair.doi.dedup.....a80ace011535ac71c55a861b195d97b2 |
| قاعدة البيانات: | OpenAIRE |
| تدمد: | 19326203 |
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