Diagnostic Molecular Pathology最新文献

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Diagnostic Molecular Pathology Pub Date : 2017-12-31 DOI: 10.1016/B978-0-12-800886-7.00053-4

引用次数: 0

Molecular Testing in Emerging Infectious Diseases 新发传染病的分子检测

Diagnostic Molecular Pathology Pub Date : 2016-10-14 DOI: 10.1016/B978-0-12-800886-7.00015-7

J. Dong, N. Ismail, D. Walker

引用次数: 3

Molecular Testing for Respiratory Viruses 呼吸道病毒的分子检测

Diagnostic Molecular Pathology Pub Date : 2016-10-14 DOI: 10.1016/B978-0-12-800886-7.00011-X

K. Stellrecht

引用次数: 8

Development and clinical validation of a multiplex real-time PCR assay for herpes simplex and varicella zoster virus. 单纯疱疹病毒和水痘带状疱疹病毒多重实时PCR检测方法的建立和临床验证。

Diagnostic Molecular Pathology Pub Date : 2013-12-01 DOI: 10.1097/PDM.0b013e3182914291

Thean Yen Tan, Hao Zou, Danny Chee Tiong Ong, Khor Jia Ker, Martin Tze Wei Chio, Rachael Yu Lin Teo, Mark Jean Aan Koh

{"title":"Development and clinical validation of a multiplex real-time PCR assay for herpes simplex and varicella zoster virus.","authors":"Thean Yen Tan, Hao Zou, Danny Chee Tiong Ong, Khor Jia Ker, Martin Tze Wei Chio, Rachael Yu Lin Teo, Mark Jean Aan Koh","doi":"10.1097/PDM.0b013e3182914291","DOIUrl":"https://doi.org/10.1097/PDM.0b013e3182914291","url":null,"abstract":"Herpes simplex virus (HSV) and varicella zoster virus (VZV) are related members of the Herpesviridae family and are well-documented human pathogens causing a spectrum of diseases, from mucocutaneous disease to infections of the central nervous system. This study was carried out to evaluate and validate the performance of a multiplex real-time polymerase chain reaction (PCR) assay in detecting and differentiating HSV1, HSV2, and VZV from clinical samples. Consensus PCR primers for HSV were designed from the UL30 component of the DNA polymerase gene of HSV, with 2 separate hydrolysis probes designed to differentiate HSV1 and HSV2. Separate primers and a probe were also designed against the DNA polymerase gene of VZV. A total of 104 clinical samples were available for testing by real-time PCR, conventional PCR, and viral culture. The sensitivity and specificity of the real-time assay was calculated by comparing the multiplex PCR result with that of a combined standard of virus culture and conventional PCR. The sensitivity of the real-time assay was 100%, with specificity ranging from 98% to 100% depending on the target gene. Both PCR methods detected more positive samples for HSV or VZV, compared with conventional virus culture. This multiplex PCR assay provides accurate and rapid diagnostic capabilities for the diagnosis and differentiation of HSV1, HSV2, and VZV infections, with the presence of an internal control to monitor for inhibition of the PCR reaction. ","PeriodicalId":11235,"journal":{"name":"Diagnostic Molecular Pathology","volume":"22 4","pages":"245-8"},"PeriodicalIF":0.0,"publicationDate":"2013-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1097/PDM.0b013e3182914291","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"31834415","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

Pyrosequencing for EGFR mutation detection: diagnostic accuracy and clinical implications. 焦磷酸测序用于EGFR突变检测:诊断准确性和临床意义。

Diagnostic Molecular Pathology Pub Date : 2013-12-01 DOI: 10.1097/PDM.0b013e3182893f55

Nora Sahnane, Rossana Gueli, Maria G Tibiletti, Barbara Bernasconi, Michele Stefanoli, Francesca Franzi, Graziella Pinotti, Carlo Capella, Daniela Furlan

{"title":"Pyrosequencing for EGFR mutation detection: diagnostic accuracy and clinical implications.","authors":"Nora Sahnane, Rossana Gueli, Maria G Tibiletti, Barbara Bernasconi, Michele Stefanoli, Francesca Franzi, Graziella Pinotti, Carlo Capella, Daniela Furlan","doi":"10.1097/PDM.0b013e3182893f55","DOIUrl":"https://doi.org/10.1097/PDM.0b013e3182893f55","url":null,"abstract":"EGFR-activating mutations predict responsiveness to EGFR tyrosine kinase inhibitors (TKIs) in non-small cell lung cancer (NSCLC) patients. Mutation screening is crucial to support therapeutic decisions and is commonly conducted using dideoxy sequencing, although its sensitivity is suboptimal in clinical settings. To evaluate the diagnostic performance of pyrosequencing and dideoxy sequencing, we examined EGFR mutation status in a retrospective cohort of 53 patients with NSCLCs clinically selected for TKI therapy and whose clinical outcome was available. Moreover, pyrosequencing quantitative results were compared with EGFR amplification data. EGFR mutations were investigated by pyrosequencing and by dideoxy sequencing. Detection rates of both methods were determined by titration assays using NCI-H1975 and HCC-827 cell lines. Increased EGFR copy number was assessed by fluorescence in situ hybridization (FISH). Pyrosequencing showed a higher detection rate than dideoxy sequencing. Tumor control rate of cases with mutant and wild-type EGFR was 86% and 29%, respectively. EGFR amplification was significantly associated with EGFR mutation and a positive correlation between high percentages of mutant alleles and clinical response to TKI was observed. We concluded that pyrosequencing is more sensitive than dideoxy sequencing in mutation screening for EGFR mutations. Detection rate of dideoxy sequencing was suboptimal when low frequencies of mutant alleles or low tumor cell contents were observed. Pyrosequencing enables quantification of mutant alleles that correlates well with increased EGFR copy number assessed by FISH. Pyrosequencing should be used in molecular diagnostic of NSCLC to appropriately select patients who are likely to benefit from TKI therapy. ","PeriodicalId":11235,"journal":{"name":"Diagnostic Molecular Pathology","volume":"22 4","pages":"196-203"},"PeriodicalIF":0.0,"publicationDate":"2013-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1097/PDM.0b013e3182893f55","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"31834412","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

A comparative study of standard cytogenetic evaluation and molecular karyotyping for products of conception. 受孕产物标准细胞遗传学评价与分子核型的比较研究。

Diagnostic Molecular Pathology Pub Date : 2013-12-01 DOI: 10.1097/PDM.0b013e31829265ab

Leanne Baxter, Nandini Adayapalam

{"title":"A comparative study of standard cytogenetic evaluation and molecular karyotyping for products of conception.","authors":"Leanne Baxter, Nandini Adayapalam","doi":"10.1097/PDM.0b013e31829265ab","DOIUrl":"https://doi.org/10.1097/PDM.0b013e31829265ab","url":null,"abstract":"Genetic analysis of fetal tissue provides valuable information regarding the underlying causes of miscarriage. However, current analysis techniques are limited and expensive. This trial compared a molecular multiplex, bead-based suspension array, KaryoLite Bacs on Beads, with conventional tissue culture and G-banded karyotype techniques. A 92% overall success rate was achieved. This method detected a cryptic deletion of a 7q subtelomeric region, a case of 25% mosaic trisomy 14, and 2 unbalanced subtelomeric rearrangements due to familial balanced translocations. Twenty (24%) of the 83 samples analyzed, that failed to yield a cytogenetic result due to culture failure, were successfully assayed using the suspension array. Genomic imbalances including trisomies and subtelomeric deletions were detected in 3 cases (15%) of previously failed cases. This method is limited by its inability to detect polyploidy, which is significant in first trimester loss. However, this can be readily overcome by prescreening using florescent in situ hybridization. Data indicates that KaryoLite BoBs molecular testing is superior to conventional cytogenetic evaluation in several key areas, including success rate (95% vs. 76%, for this study group), cost, turnaround time (2 vs. up to 28 d), and subjective result interpretation. ","PeriodicalId":11235,"journal":{"name":"Diagnostic Molecular Pathology","volume":"22 4","pages":"228-35"},"PeriodicalIF":0.0,"publicationDate":"2013-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1097/PDM.0b013e31829265ab","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"31834416","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

An analytical method for the quantification of hERG1 channel gene expression in human colorectal cancer. 人结直肠癌中hERG1通道基因表达的定量分析方法。

Diagnostic Molecular Pathology Pub Date : 2013-12-01 DOI: 10.1097/PDM.0b013e31828e55c7

Angelo Fortunato, Luca Gasparoli, Sara Falsini, Luca Boni, Boni Luca, Annarosa Arcangeli

{"title":"An analytical method for the quantification of hERG1 channel gene expression in human colorectal cancer.","authors":"Angelo Fortunato, Luca Gasparoli, Sara Falsini, Luca Boni, Boni Luca, Annarosa Arcangeli","doi":"10.1097/PDM.0b013e31828e55c7","DOIUrl":"https://doi.org/10.1097/PDM.0b013e31828e55c7","url":null,"abstract":"Cancer molecular investigation revealed a huge molecular heterogeneity between different types of cancers as well as among cancer patients affected by the same cancer type. This implies the necessity of a personalized approach for cancer diagnosis and therapy, on the basis of the development of standardized protocols to facilitate the application of molecular techniques in the clinical decision-making process. Ion channels encoding genes are acquiring increasing relevance in oncological translational studies, representing new candidates for molecular diagnostic and therapeutic purposes. Hence, the development of molecular protocols for the quantification of ion channels encoding genes in tumor specimens may have relevance for diagnostic and prognostic investigation. Two main hindrances must be overcome for these purposes: the use of formalin-fixed and paraffin-embedded samples for gene expression analysis and the physiological expression of ion channels in excitable cells, potentially present in the tumor sample. We here propose a method for hERG1 gene quantification in colorectal cancer samples in both cryopreserved and formalin-fixed and paraffin-embedded samples. An analytical method was developed to estimate hERG1 gene expression exclusively in epithelial cancer cells. Indeed, we found that the hERG1 gene was expressed at significant levels by myofibroblasts present in the tumor stroma. This method was based on the normalization on a smooth muscle-myofibroblast-specific gene, MYH11, with no need of microdissection. By applying this method, hERG1 expression turned out to correlate with VEGF-A expression, confirming previous immunohistochemical data. ","PeriodicalId":11235,"journal":{"name":"Diagnostic Molecular Pathology","volume":"22 4","pages":"215-21"},"PeriodicalIF":0.0,"publicationDate":"2013-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1097/PDM.0b013e31828e55c7","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"31834413","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: 4

EGFR autophosphorylation but not protein score correlates with histologic and molecular subtypes in lung adenocarcinoma. EGFR自磷酸化与肺腺癌的组织学和分子亚型相关，但与蛋白评分无关。

Diagnostic Molecular Pathology Pub Date : 2013-12-01 DOI: 10.1097/PDM.0b013e3182936957

Judit Moldvay, Tamás Barbai, Krisztina Bogos, Violetta Piurko, János Fillinger, Helmut H Popper, József Tímár

{"title":"EGFR autophosphorylation but not protein score correlates with histologic and molecular subtypes in lung adenocarcinoma.","authors":"Judit Moldvay, Tamás Barbai, Krisztina Bogos, Violetta Piurko, János Fillinger, Helmut H Popper, József Tímár","doi":"10.1097/PDM.0b013e3182936957","DOIUrl":"https://doi.org/10.1097/PDM.0b013e3182936957","url":null,"abstract":"Established clinicopathologic characteristics of non-small cell lung cancer patients define a subgroup responding better to EGFR-TK inhibitors: adenocarcinoma histology, ethnicity, sex, smoking status, presence of activating EGFR mutation, and/or K-RAS wild type. However, EGFR mutation does not automatically lead to increased activity of the protein influenced by several factors. As adenocarcinoma can be further divided into histologic subclasses, we compared adenocarcinomas without lepidic growth pattern (NLAC) to those characterized by pure or predominant lepidic growth (LAC) for EGFR protein expression and autophosphorylation activity (Y1173), as determined by immunohistochemistry. This pretarget therapy cohort comprised a total of 110 surgically operated patients of stage I non-small cell lung cancer: 49 NLAC and 61 LAC variants. The LAC group had a significantly better prognosis and the incidence of phospho-EGFR-positive tumors was significantly higher compared with NLAC. Patient sex did not influence EGFR activity, but the incidence of pEGFR-positive tumors was significantly lower among smoker patients. There was no statistically significant difference in EGFR or KRAS mutation frequencies between the 2 groups. In NLAC, pEGFR-positive tumors occurred exclusively among EGFR-mutant/K-RAS wild-type tumors. On the contrary, in LAC tumors, pEGFR-positive tumors were similarly frequent in the EGFR or K-RAS mutant groups indicating an interesting feedback activation of EGFR signaling in K-RAS mutant tumors. Our data also indicate that EGFR mutation leads to EGFR autophosphorylation only in a small fraction of adenocarcinoma patients, which might have clinical significance. ","PeriodicalId":11235,"journal":{"name":"Diagnostic Molecular Pathology","volume":"22 4","pages":"204-9"},"PeriodicalIF":0.0,"publicationDate":"2013-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1097/PDM.0b013e3182936957","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"31834418","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: 2

Influence of parasite density and sample storage time on the reliability of Entamoeba histolytica-specific PCR from formalin-fixed and paraffin-embedded tissues. 寄生虫密度和样品保存时间对福尔马林固定和石蜡包埋组织中溶组织内阿米巴特异性PCR可靠性的影响。

Diagnostic Molecular Pathology Pub Date : 2013-12-01 DOI: 10.1097/PDM.0b013e3182936936

Hagen Frickmann, Klara Tenner-Racz, Petra Eggert, Norbert G Schwarz, Sven Poppert, Egbert Tannich, Ralf M Hagen

{"title":"Influence of parasite density and sample storage time on the reliability of Entamoeba histolytica-specific PCR from formalin-fixed and paraffin-embedded tissues.","authors":"Hagen Frickmann, Klara Tenner-Racz, Petra Eggert, Norbert G Schwarz, Sven Poppert, Egbert Tannich, Ralf M Hagen","doi":"10.1097/PDM.0b013e3182936936","DOIUrl":"https://doi.org/10.1097/PDM.0b013e3182936936","url":null,"abstract":"We report on the reliability of polymerase chain reaction (PCR) for the detection of Entamoeba histolytica from formalin-fixed, paraffin-embedded tissue in comparison with microscopy and have determined predictors that may influence PCR results. E. histolytica-specific and Entamoeba dispar-specific real-time PCR and microscopy from adjacent histologic sections were performed using a collection of formalin-fixed, paraffin-embedded tissue specimens obtained from patients with invasive amebiasis. Specimens had been collected during the previous 4 decades. Association of sample age, parasite density, and reliability of PCR was analyzed. E. histolytica PCR was positive in 20 of 34 biopsies (58.8%); 2 of these 20 were microscopically negative for amebae in neighboring tissue sections. PCR was negative in 9 samples with visible amebae in neighboring sections and in 5 samples without visible parasites in neighboring sections. PCR was negative in all specimens that were older than 3 decades. Low parasite counts and sample ages older than 20 years were predictors for false-negative PCR results. All samples were negative for E. dispar DNA. PCR is suitable for the detection of E. histolytica in formalin-fixed, paraffin-embedded tissue samples that are younger than 2 decades and that contain intermediate to high parasite numbers. Negative results in older samples were due to progressive degradation of DNA over time as indicated by control PCRs targeting the human 18S rRNA gene. Moreover, our findings support previous suggestions that only E. histolytica but not E. dispar is responsible for invasive amebiasis.","PeriodicalId":11235,"journal":{"name":"Diagnostic Molecular Pathology","volume":"22 4","pages":"236-44"},"PeriodicalIF":0.0,"publicationDate":"2013-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1097/PDM.0b013e3182936936","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"31834417","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: 20

Methylation profile analysis of DNA repair genes in hepatocellular carcinoma with MS-MLPA. 肝癌DNA修复基因甲基化谱的MS-MLPA分析。

Diagnostic Molecular Pathology Pub Date : 2013-12-01 DOI: 10.1097/PDM.0b013e31828ed856

Ozge Ozer, Banu Bilezikci, Sema Aktas, Feride I Sahin

{"title":"Methylation profile analysis of DNA repair genes in hepatocellular carcinoma with MS-MLPA.","authors":"Ozge Ozer, Banu Bilezikci, Sema Aktas, Feride I Sahin","doi":"10.1097/PDM.0b013e31828ed856","DOIUrl":"https://doi.org/10.1097/PDM.0b013e31828ed856","url":null,"abstract":"Hepatocellular carcinoma (HCC) is one of the rare tumors with well-defined risk factors. The multifactorial etiology of HCC can be explained by its complex molecular pathogenesis. In the current study, the methylation status of 7 genes involved in DNA repair mechanisms, namely MLH1, PMS2, MSH6, MSH2, MGMT, MSH3, and MLH3, was investigated in tumor samples from HCC patients, using the methylation-specific-multiplex ligated probe amplification method and the results were correlated with available clinical findings. The most common etiological factor in these cases was the presence of hepatitis B alone (47.2%). Among the 56 cases that were studied, promoter methylation was detected in at least one of the genes in 27 (48.2%) cases, only in 1 gene in 13 (23.2%) cases, and in >1 gene in 14 (25%) cases. Of the 7 genes investigated, methylation was most frequently observed in MSH3, in 14 (25%) cases. Methylation of at least 1 gene was significantly more frequent in patients with single tumors than multifocal tumors. There were significant differences regarding hepatitis B status, Child Class, tumor number, grade, and TNM stage in cases where PMS2 methylation was detected. Our results suggest that methylation of genes involved in mismatch repair may be responsible in the pathogenesis of HCC, and evaluating changes in multiple genes in these pathways simultaneously would be more informative. Despite being a robust and relatively inexpensive method, the methylation-specific-multiplex ligated probe amplification assay could be more extensively applied with improvements in the currently intricate data analysis component. ","PeriodicalId":11235,"journal":{"name":"Diagnostic Molecular Pathology","volume":"22 4","pages":"222-7"},"PeriodicalIF":0.0,"publicationDate":"2013-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1097/PDM.0b013e31828ed856","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"31834414","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: 3