Diagnostic Molecular Pathology最新文献_第4页

Effect of HPV assay choice on perceived prevalence in a population-based sample. HPV检测选择对基于人群样本的感知患病率的影响。

Diagnostic Molecular Pathology Pub Date : 2013-06-01 DOI: 10.1097/PDM.0b013e31827f3f7e

Kate Cuschieri, Kim Kavanagh, Katy Sinka, Chris Robertson, Heather Cubie, Catherine Moore, Martin Donaghy

{"title":"Effect of HPV assay choice on perceived prevalence in a population-based sample.","authors":"Kate Cuschieri, Kim Kavanagh, Katy Sinka, Chris Robertson, Heather Cubie, Catherine Moore, Martin Donaghy","doi":"10.1097/PDM.0b013e31827f3f7e","DOIUrl":"https://doi.org/10.1097/PDM.0b013e31827f3f7e","url":null,"abstract":"Human papillomavirus (HPV) immunization programs clearly have considerable potential to reduce HPV-associated disease; they are also resource-intense; so, it is essential that their effectiveness is determined accurately and in a timely way. Measuring circulating HPV types in a population can provide an early measure of vaccine impact. We assessed the impact of HPV assay on the observed population prevalence of HPV in women who provided samples as part of a National HPV Immunisation Surveillance Exercise. A total of 1145 liquid-based cytology samples, 326 self-taken swabs, and 371 urine samples were tested with a line-blot assay (the Digene reverse hybridization HPV genotyping assay) and a luminex-based assay (the Mulitmetrix HPV genotyping assay). Assay agreement was determined for the different sample types. Positivity (according to assay) was compared at different levels ranging from positive for HPV 16 and/or 18 to positive for any one of the 18 HPV types common to both assays. The luminex assay consistently detected a higher prevalence of HPV--up to 10% for HPV types common to both assays. In addition, disagreement for HPV 16 and/or 18 was observed in around 9% of the overall sample, with an associated κ score of 0.74. These data indicate that assay choice has a significant impact on observed prevalence of HPV, including vaccine types. The impact of any change of assay during longitudinal surveillance programs should thus be taken into account to avoid confounding the assessment of any vaccine-induced changes.","PeriodicalId":11235,"journal":{"name":"Diagnostic Molecular Pathology","volume":"22 2","pages":"85-90"},"PeriodicalIF":0.0,"publicationDate":"2013-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1097/PDM.0b013e31827f3f7e","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"31394142","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 19

Analytical validation of a practical molecular assay prognostic of survival in nonsquamous non-small cell lung cancer. 一种实用的非鳞状非小细胞肺癌生存预后分子测定方法的分析验证。

Diagnostic Molecular Pathology Pub Date : 2013-06-01 DOI: 10.1097/PDM.0b013e318273fb61

Johannes R Kratz, Patrick T Tham, Michael S Mulvihill, Fatemeh Ziaei, Mahashweta Roshni Ray, Jerry W Hurst, Mark R Segal, David M Berryman, Wenjiang Chu, Biao He, David M Jablons, Michael J Mann

{"title":"Analytical validation of a practical molecular assay prognostic of survival in nonsquamous non-small cell lung cancer.","authors":"Johannes R Kratz, Patrick T Tham, Michael S Mulvihill, Fatemeh Ziaei, Mahashweta Roshni Ray, Jerry W Hurst, Mark R Segal, David M Berryman, Wenjiang Chu, Biao He, David M Jablons, Michael J Mann","doi":"10.1097/PDM.0b013e318273fb61","DOIUrl":"https://doi.org/10.1097/PDM.0b013e318273fb61","url":null,"abstract":"A molecular assay prognostic of survival in resected nonsquamous non-small cell lung cancer designed to meet the need for improved risk stratification in early-stage disease has recently been described. This assay measures the expression levels of 14 genes using RNA extracted from formalin-fixed, paraffin-embedded (FFPE) tissues. The assay underwent blinded clinical validation in 2 large international cohorts involving approximately 1500 patients; the analytical precision and reproducibility of this assay, however, have not yet been reported. For each of the 14 TaqMan quantitative polymerase chain reaction (PCR) primer and probe sets used in the molecular prognostic assay, the linear range, PCR efficiency, limits of blank, limits of quantitation, and quantitative bias were determined using serial dilutions of pooled RNA extracted from FFPE samples. The reproducibility of the entire molecular assay was determined by performing repeat testing of FFPE samples over multiple days. The linear range of individual quantitative TaqMan PCR primer and probe sets was between 2(10)- and 2(15)-fold input RNA. The median C(T) of the quantitative PCR primer and probe sets at 10 ng of input RNA was 24.3; the median efficiency was 91.2%. The median quantitative bias across all quantitative PCR primer and probe sets was 0.75% (range, 0.32% to 1.32%). In repeat testing, the mean SD of the risk score (scaled from 1 to 100) was 2.18, with a mean coefficient of variation of 0.08. The molecular prognostic assay presented in this study demonstrates high precision and reproducibility, validating its clinical utility as a reliable prognostic tool that can contribute to the management of patients with early-stage disease.","PeriodicalId":11235,"journal":{"name":"Diagnostic Molecular Pathology","volume":"22 2","pages":"65-9"},"PeriodicalIF":0.0,"publicationDate":"2013-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1097/PDM.0b013e318273fb61","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"31394139","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 19

Xp11.2 translocation renal cell carcinoma with PSF-TFE3 rearrangement. Xp11.2易位性肾细胞癌伴PSF-TFE3重排。

Diagnostic Molecular Pathology Pub Date : 2013-06-01 DOI: 10.1097/PDM.0b013e318278962e

Minghao Zhong, Paul Weisman, Bing Zhu, Maria Brassesco, Youfeng Yang, W Marston Linehan, Maria J Merino, David Zhang, Stephen Rohan, Dongming Cai, Ximing Yang

{"title":"Xp11.2 translocation renal cell carcinoma with PSF-TFE3 rearrangement.","authors":"Minghao Zhong, Paul Weisman, Bing Zhu, Maria Brassesco, Youfeng Yang, W Marston Linehan, Maria J Merino, David Zhang, Stephen Rohan, Dongming Cai, Ximing Yang","doi":"10.1097/PDM.0b013e318278962e","DOIUrl":"https://doi.org/10.1097/PDM.0b013e318278962e","url":null,"abstract":"Xp11.2 translocation renal cell carcinoma (Xp11.2 RCC) is a subtype of RCC characterized by translocations involving a breakpoint at the TFE3 gene (Xp11.2). Moderate to strong nuclear TFE3 immunoreactivity has been recognized as a specific diagnostic marker for this type of tumor. However, exclusive cytoplasmic localization of a TFE3 fusion protein was reported in UOK 145 cells, a cell line derived from an Xp11.2 RCC harboring the PSF-TFE3 translocation. If reproducible using immunohistochemistry (IHC), this finding would have important implications for pathologists in the diagnosis of Xp11.2 RCC, calling into question the specificity of nuclear immunoreactivity for TFE3 in these tumors. The purpose of this study was to determine whether the above-noted cytoplasmic localization of the TFE3 fusion protein could be reproduced using IHC. UOK 145 cells and fresh frozen tissue from 2 clinical cases of Xp11.2 RCC found to harbor the PSF-TFE3 gene rearrangement (by cytogenetic testing) were collected. All samples were subjected to histopathologic evaluation by board-certified pathologists, TFE3 IHC, reverse transcription polymerase chain reaction, and Sanger sequencing analysis. A strong nuclear TFE3 immunoreactivity was demonstrated in all samples including the UOK 145 cell line. No cytoplasmic immunoreactivity was seen. Reverse transcription polymerase chain reaction and Sanger sequencing confirmed the previously reported PSF-TFE3 gene fusion between exon 9 of PSF and exon 6 of TFE3 in the UOK 145 cell line and in one of 2 clinical cases of Xp11.2 RCC. A novel PSF-TFE3 gene fusion between exon 9 of PSF and exon 5 of TFE3 was detected in the second clinical case of Xp11.2 RCC.","PeriodicalId":11235,"journal":{"name":"Diagnostic Molecular Pathology","volume":"22 2","pages":"107-11"},"PeriodicalIF":0.0,"publicationDate":"2013-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1097/PDM.0b013e318278962e","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"31395121","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 6

Expression analysis on archival material revisited: isolation and quantification of RNA extracted from FFPE samples. 档案材料的表达分析:从FFPE样品中提取的RNA的分离和定量。

Diagnostic Molecular Pathology Pub Date : 2013-03-01 DOI: 10.1097/PDM.0b013e318269de3b

Christophe Deben, Karen Zwaenepoel, Carolien Boeckx, An Wouters, Patrick Pauwels, Marc Peeters, Filip Lardon, Marc Baay, Vanessa Deschoolmeester

{"title":"Expression analysis on archival material revisited: isolation and quantification of RNA extracted from FFPE samples.","authors":"Christophe Deben, Karen Zwaenepoel, Carolien Boeckx, An Wouters, Patrick Pauwels, Marc Peeters, Filip Lardon, Marc Baay, Vanessa Deschoolmeester","doi":"10.1097/PDM.0b013e318269de3b","DOIUrl":"https://doi.org/10.1097/PDM.0b013e318269de3b","url":null,"abstract":"Background: Formalin-fixed paraffin-embedded (FFPE) tissue is the most readily available source of RNA for the gene expression studies. The main disadvantage is the poor quality of isolated RNA. Our group recently compared 5 commercially available RNA isolation kits and concluded that the RNeasy FFPE kit from Qiagen was the most appropriate one. However, this kit has been discontinued and replaced by a new version. In this study both kits were compared, and spectrophotometric and fluorometric analyses for quantification of RNA samples extracted from FFPE tissue.Methods: Both RNeasy FFPE kits were compared for the total RNA and DNA yields, purity, and raw cycle threshold. Quantity and quality of the isolated RNA was measured using the NanoDrop ND-1000 spectrophotometer and Qubit 2.0 fluorometer.Results: The average concentration of RNA extracted from FFPE tissue measured using the NanoDrop was 32.0%±9.5% higher than the concentration measured using the Qubit. When measuring an RNA sample extracted from a cell line, the concentration measured using both methods was similar. When comparing both RNeasy FFPE kits, marginal differences were observed for total RNA yield, purity, and raw cycle threshold. However, the residual DNA in the samples isolated using the old kit was higher than in the samples isolated using the new kit.Conclusions: A fluorometric analysis is more suitable for quantification of RNA samples extracted from FFPE tissue compared with spectrophotometric analysis. For RNA isolation from FFPE tissue, both old and new RNeasy FFPE kits were adequate. The new kit resulted in more efficient DNA removal.","PeriodicalId":11235,"journal":{"name":"Diagnostic Molecular Pathology","volume":"22 1","pages":"59-64"},"PeriodicalIF":0.0,"publicationDate":"2013-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1097/PDM.0b013e318269de3b","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"31204831","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 27

Detection of minor clones with internal tandem duplication mutations of FLT3 gene in acute myeloid leukemia using delta-PCR. 应用delta-PCR检测急性髓系白血病FLT3基因内串联重复突变的小克隆。

Diagnostic Molecular Pathology Pub Date : 2013-03-01 DOI: 10.1097/PDM.0b013e31825d81f4

Katie Beierl, Li-Hui Tseng, Russell Beierl, Lisa Haley, Christopher D Gocke, James R Eshleman, Ming-Tseh Lin

{"title":"Detection of minor clones with internal tandem duplication mutations of FLT3 gene in acute myeloid leukemia using delta-PCR.","authors":"Katie Beierl, Li-Hui Tseng, Russell Beierl, Lisa Haley, Christopher D Gocke, James R Eshleman, Ming-Tseh Lin","doi":"10.1097/PDM.0b013e31825d81f4","DOIUrl":"https://doi.org/10.1097/PDM.0b013e31825d81f4","url":null,"abstract":"Internal tandem duplication (ITD) mutations of the FLT3 gene have been associated with inferior prognosis of acute myeloid leukemia. Detection of minor clones or minimal residual clones with ITD mutations is desirable, but is challenging when the mutant signal determined by polymerase chain reaction (PCR) and capillary electrophoresis is weak. In this study, we applied delta-PCR, which is a triple-primer strategy, to ensure PCR specificity and improve the sensitivity to 0.1% leukemic cells with ITD mutation. We also applied a reference peak to calculate ITD allelic burdens of <2% threshold of technical limitation for evaluating the relative ratio of 2 signals by capillary electrophoresis. Delta-PCR was able to detect single or multiple ITD mutations with an allelic burden (peak height ratio of mutant allele and wild-type allele) ranging from 0.4% to >100% among all 31 cases with previous documented ITD mutations. In one of the 3 cases with previously reported negative ITD mutation in the initial diagnostic specimen and ITD-positive results in the follow-up specimens, an ITD of 0.04% allele burden was retrospectively detected in the initial diagnosis specimen using delta-PCR. We also demonstrated that minor ITD mutant clones with an allelic burden of <1% present at diagnosis may become a dominant clone at the later refractory status, suggesting that detection of leukemic clones with allelic burdens of <1% may be clinically significant. Delta-PCR can detect ITD mutations with improved sensitivity and specificity and may be useful for the detection of minimal residual leukemia.","PeriodicalId":11235,"journal":{"name":"Diagnostic Molecular Pathology","volume":"22 1","pages":"1-9"},"PeriodicalIF":0.0,"publicationDate":"2013-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1097/PDM.0b013e31825d81f4","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"31204827","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 12

Identification of uncommon PIK3CA mutations in lung cancer by using pyrosequencing. 利用焦磷酸测序技术鉴定肺癌中罕见的PIK3CA突变。

Diagnostic Molecular Pathology Pub Date : 2013-03-01 DOI: 10.1097/PDM.0b013e31825f5f93

Verena Schildgen, Jessica Lüsebrink, Jan D Appel, Christine Wübben, Walburga Engel-Riedel, Corinna Ludwig, Erich Stoelben, Oliver Schildgen, Michael Brockmann

{"title":"Identification of uncommon PIK3CA mutations in lung cancer by using pyrosequencing.","authors":"Verena Schildgen, Jessica Lüsebrink, Jan D Appel, Christine Wübben, Walburga Engel-Riedel, Corinna Ludwig, Erich Stoelben, Oliver Schildgen, Michael Brockmann","doi":"10.1097/PDM.0b013e31825f5f93","DOIUrl":"https://doi.org/10.1097/PDM.0b013e31825f5f93","url":null,"abstract":"Introduction: Phospatidylinositol-3-kinases (PI3K) play an important role in various cell processes. Oncogenic mutations in the PIK3CA gene, which codes for the catalytic subunit, have been identified in various malignancies and activate the PI3K/AKT/mTOR pathway, which is a critical driver of tumorigenesis.Methods: We tested 41 tumor samples with known KRAS, BRAF, and EGFR mutation status for the occurrence of mutations in the PIK3CA gene, using a pyrosequencing assay.Results: Pyrosequencing revealed 2 mutations (4.9%) in the PIK3CA gene, one in exon 9 and the other in exon 20. Both mutations have not been identified yet in lung tumor tissue.Discussion: The screening of our small patient cohort by pyrosequencing identified 2 mutations (4.9%) in PIK3CA, one in exon 9 (Q546H) and the other in exon 20 (M1043T). Both mutations have not been described in lung tumors yet and seem to be rather uncommon mutations. Future screening of large patient cohorts with pyrosequencing may contribute to the detection of more mutations in lung cancer because of the low limit of detections of this method and may contribute to a better understanding of the interaction of mutations and tumorigenesis.","PeriodicalId":11235,"journal":{"name":"Diagnostic Molecular Pathology","volume":"22 1","pages":"22-7"},"PeriodicalIF":0.0,"publicationDate":"2013-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1097/PDM.0b013e31825f5f93","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"31204829","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 9

Deparaffinization and lysis by hydrothermal pressure (pressure cooking) coupled with chaotropic salt column purification: a rapid and efficient method of DNA extraction from formalin-fixed paraffin-embedded tissue. 水热加压(压力蒸煮)脱蜡裂解-朝向盐柱纯化:一种快速高效的福尔马林固定石蜡包埋组织DNA提取方法。

Diagnostic Molecular Pathology Pub Date : 2013-03-01 DOI: 10.1097/PDM.0b013e318263f092

Haohao Zhong, Yan Liu, Monica Talmor, Bingquan Wu, Pei Hui

{"title":"Deparaffinization and lysis by hydrothermal pressure (pressure cooking) coupled with chaotropic salt column purification: a rapid and efficient method of DNA extraction from formalin-fixed paraffin-embedded tissue.","authors":"Haohao Zhong, Yan Liu, Monica Talmor, Bingquan Wu, Pei Hui","doi":"10.1097/PDM.0b013e318263f092","DOIUrl":"https://doi.org/10.1097/PDM.0b013e318263f092","url":null,"abstract":"We report a hydrothermal pressure method (pressure cooking) for simultaneous deparaffinization and lysis of formalin-fixed paraffin-embedded tissue followed by conventional chaotropic salt column purification to obtain high-quality DNA. Using this method, the release of DNA occurred within the first minute of treatment, reaching the maximum at 5 minutes. An optimal treatment window was between 5 and 30 minutes. The extracted DNA was of high quality as determined by the 260/280 absorbance ratios, and the quantity of DNA extracted was linear with the input tissue amount. In paired sample experiments (N=19), the quantity of DNA extracted by hydrothermal pressure treatment was comparable to that obtained through the conventional xylene deparaffinization and protease K digestion method. The integrity of the recovered DNA was also comparable, evidenced by polymerase chain reaction amplifications of variable-sized amplicons in tissue samples archived from 0.2 to 22 years (N=14). The high quality of DNA was further confirmed by polymerase chain reaction amplification and Sanger sequencing analysis of representative exons of the EGFR gene in human non-small cell lung cancer tissue samples. In summary, this novel method offers DNA release from formalin-fixed paraffin-embedded tissue with unprecedented simplicity, speed, biohazard safety, and cost-efficiency. Combined with chaotropic salt column purification, high-quality DNA can be prepared for downstream applications in <30 minutes.","PeriodicalId":11235,"journal":{"name":"Diagnostic Molecular Pathology","volume":"22 1","pages":"52-8"},"PeriodicalIF":0.0,"publicationDate":"2013-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1097/PDM.0b013e318263f092","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"31204830","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 21

A novel COLD-PCR/FMCA assay enhances the detection of low-abundance IDH1 mutations in gliomas. 一种新的COLD-PCR/FMCA检测增强了对胶质瘤中低丰度IDH1突变的检测。

Diagnostic Molecular Pathology Pub Date : 2013-03-01 DOI: 10.1097/PDM.0b013e31826c7ff8

Brendan Pang, Mary B Durso, Ronald L Hamilton, Marina N Nikiforova

{"title":"A novel COLD-PCR/FMCA assay enhances the detection of low-abundance IDH1 mutations in gliomas.","authors":"Brendan Pang, Mary B Durso, Ronald L Hamilton, Marina N Nikiforova","doi":"10.1097/PDM.0b013e31826c7ff8","DOIUrl":"https://doi.org/10.1097/PDM.0b013e31826c7ff8","url":null,"abstract":"Point mutations in isocitrate dehydrogenase 1 (IDH1) have been identified in many gliomas. The detection of IDH1 mutations becomes challenging on suboptimal glioma biopsies when a limited number of tumor cells is available for analysis. Coamplification at lower denaturing-polymerase chain reaction (COLD-PCR) is a PCR technique that deliberately lowers the denaturing cycle temperature to selectively favor amplification of mutant alleles, allowing for the sensitive detection of low-abundance mutations. We developed a novel COLD-PCR assay on the LightCycler platform (Roche, Applied Science, Indianapolis, IN), using post-PCR fluorescent melting curve analysis (FMCA) for the detection of mutant IDH1 with a detection limit of 1%. Thirty-five WHO grade I to IV gliomas and 9 non-neoplastic brain and spinal cord biopsies were analyzed with this technique and the results were compared with the conventional real-time PCR and the Sanger sequencing analysis. COLD-PCR/FMCA was able to detect the most common IDH1 R132H mutation and rare mutation types including R132H, R132C, R132L, R132S, and R132G mutations. Twenty-five glioma cases were positive for IDH1 mutations by COLD-PCR/FMCA, and 23 gliomas were positive by the conventional real-time PCR and Sanger sequencing. A pilocytic astrocytoma (PA I) and a glioblastoma multiforme (GBM IV) showed low-abundance IDH1 mutations detected by COLD-PCR/FMCA. The remaining 10 glioma and 9 non-neoplastic samples were negative by all the 3 methods. In summary, we report a novel COLD-PCR/FMCA method that provides rapid and sensitive detection of IDH1 mutations in formalin-fixed paraffin-embedded tissue and can be used in the clinical setting to assess the small brain biopsies.","PeriodicalId":11235,"journal":{"name":"Diagnostic Molecular Pathology","volume":"22 1","pages":"28-34"},"PeriodicalIF":0.0,"publicationDate":"2013-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1097/PDM.0b013e31826c7ff8","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"31204833","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 15

BRAF V600E Mutations in Endometrial Adenocarcinoma. 子宫内膜腺癌中的BRAF V600E突变。

Diagnostic Molecular Pathology Pub Date : 2013-03-01 DOI: 10.1097/PDM.0b013e31826c7fe0

Mai He, Virginia Breese, Steven Hang, Cunxian Zhang, Jinjun Xiong, Cynthia Jackson

{"title":"BRAF V600E Mutations in Endometrial Adenocarcinoma.","authors":"Mai He, Virginia Breese, Steven Hang, Cunxian Zhang, Jinjun Xiong, Cynthia Jackson","doi":"10.1097/PDM.0b013e31826c7fe0","DOIUrl":"https://doi.org/10.1097/PDM.0b013e31826c7fe0","url":null,"abstract":"The BRAF V600E somatic mutation is recognized as an oncogenic driver of many human cancers involving the MAPK/ERK pathway. Colorectal and lung cancers associated with the BRAF V600E mutation often demonstrated mucinous morphology. This study hypothesized that the BRAF V600E mutation may be associated with mucinous morphology in endometrial cancer and aimed to investigate its prevalence in mucinous (endometrial) carcinoma (MC) and endometrioid adenocarcinoma with significant mucinous differentiations (ECMD) (>10% neoplastic cells). Twenty-eight cases of endometrial cancer were selected, including 17 (60.7%) cases of MC or ECMD. All patients were Caucasian with age ranging from 50 to 87 years old (median 65). Genomic DNA was extracted from formalin-fixed paraffin-embedded tumor tissue and subjected to both real-time mutant allele-specific amplification polymerase chain reaction (PCR) and PCR amplification, followed by direct sequencing. Three (3/28, 10.7%) BRAF V600E mutations were detected by real-time mutant allele-specific amplification PCR and confirmed by direct sequencing. Two of 3 cases positive for BRAF V600E mutation were ECMDs with \"surface epithelial changes.\" KRAS mutations were found in 9 cases (32.1%), none with BRAF mutation. This is the first report of BRAF V600E mutation in endometrial cancer, indicating that it may contribute to tumorigenesis of endometrial cancer, although at a low frequency compared with KRAS mutations.","PeriodicalId":11235,"journal":{"name":"Diagnostic Molecular Pathology","volume":"22 1","pages":"35-40"},"PeriodicalIF":0.0,"publicationDate":"2013-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1097/PDM.0b013e31826c7fe0","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"31204832","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 10

Modified array-based comparative genomic hybridization detects cryptic and variant PML-RARA rearrangements in acute promyelocytic leukemia lacking classic translocations. 改良的基于阵列的比较基因组杂交检测急性早幼粒细胞白血病中缺乏经典易位的隐性和变异PML-RARA重排。

Diagnostic Molecular Pathology Pub Date : 2013-03-01 DOI: 10.1097/PDM.0b013e31825b8326

Aaron M Gruver, Heesun J Rogers, James R Cook, Blake C Ballif, Roger A Schultz, Jacqueline R Batanian, Mark J Fesler, Raymond R Tubbs

{"title":"Modified array-based comparative genomic hybridization detects cryptic and variant PML-RARA rearrangements in acute promyelocytic leukemia lacking classic translocations.","authors":"Aaron M Gruver, Heesun J Rogers, James R Cook, Blake C Ballif, Roger A Schultz, Jacqueline R Batanian, Mark J Fesler, Raymond R Tubbs","doi":"10.1097/PDM.0b013e31825b8326","DOIUrl":"https://doi.org/10.1097/PDM.0b013e31825b8326","url":null,"abstract":"Acute promyelocytic leukemia (APL) is typically defined at the molecular level by a reciprocal translocation of the promyelocytic leukemia (PML) and retinoic acid receptor α (RARA) genes. An accurate diagnosis of APL is critical for appropriate choice of therapy and prognostic assessment. Cryptic and variant rearrangements in APL are discoverable by a variety of molecular methods including fluorescence in situ hybridization (FISH), reverse transcriptase polymerase chain reaction, or gene sequencing. Rare reports of FISH-negative APL harboring cryptic rearrangements of PML-RARA detected by reverse transcriptase polymerase chain reaction or sequencing have been described. Here, we describe the detection of cryptic or variant PML-RARA rearrangements by translocation-based comparative genomic hybridization (tCGH), a recently described modification of traditional CGH technology that facilitates the detection of balanced translocations by means of the linear amplification of a potential translocation breakpoint region(s), in 2 unusual cases of APL. One tumor lacked detectable t(15;17) by karyotype and FISH, and the other tumor lacked the typical morphologic and immunophenotypic features of APL and had a variant 3-way translocation involving PML and RARA. PML-RARA translocations were identified by tCGH in both cases providing confirmation of the diagnosis of APL. These data emphasize the benefit of using complementary molecular methods including tCGH for detecting cryptic and variant PML-RARA translocations in unusual cases of APL.","PeriodicalId":11235,"journal":{"name":"Diagnostic Molecular Pathology","volume":"22 1","pages":"10-21"},"PeriodicalIF":0.0,"publicationDate":"2013-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1097/PDM.0b013e31825b8326","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"31204826","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 10