scholarly journals Quantification of Plasma miRNAs by Digital PCR for Cancer Diagnosis

2013 ◽  
Vol 8 ◽  
pp. BMI.S13154 ◽  
Author(s):  
Jie Ma ◽  
Ning Li ◽  
Maria Guarnera ◽  
Feng Jiang
Keyword(s):  
Lung Cancer ◽  
Cancer Patients ◽  
Cancer Diagnosis ◽  
Copy Number ◽  
Dynamic Range ◽  
Quantitative Polymerase Chain Reaction ◽  
Digital Pcr ◽  
Lung Cancer Patients ◽  
Plasma Microrna ◽  
Plasma Mirnas

Analysis of plasma microRNAs (miRNAs) by quantitative polymerase chain reaction (qPCR) provides a potential approach for cancer diagnosis. However, absolutely quantifying low abundant plasma miRNAs is challenging with qPCR. Digital PCR offers a unique means for assessment of nucleic acids presenting at low levels in plasma. This study aimed to evaluate the efficacy of digital PCR for quantification of plasma miRNAs and the potential utility of this technique for cancer diagnosis. We used digital PCR to quantify the copy number of plasma microRNA-21-5p (miR-21–5p) and microRNA-335–3p (miR-335–3p) in 36 lung cancer patients and 38 controls. Digital PCR showed a high degree of linearity and quantitative correlation with miRNAs in a dynamic range from 1 to 10,000 copies/μL of input, with high reproducibility. qPCR exhibited a dynamic range from 100 to 1X107 copies/μL of input. Digital PCR had a higher sensitivity to detect copy number of the miRNAs compared with qPCR. In plasma, digital PCR could detect copy number of both miR-21–5p and miR-335–3p, whereas qPCR was only able to assess miR-21–5p. Quantification of the plasma miRNAs by digital PCR provided 71.8% sensitivity and 80.6% specificity in distinguishing lung cancer patients from cancer-free subjects.

scholarly journals Digital PCR for the Analysis of MYC Copy Number Variation in Lung Cancer

2020 ◽  
Vol 2020 ◽  
pp. 1-8
Author(s):  
Alexander Brik ◽  
Daniel G. Weber ◽  
Swaantje Casjens ◽  
Peter Rozynek ◽  
Swetlana Meier ◽  
...  
Keyword(s):  
Lung Cancer ◽  
Cancer Patients ◽  
Copy Number ◽  
Digital Pcr ◽  
Copy Number Variations ◽  
Clinical Diagnostics ◽  
Gene Copy ◽  
Clinicopathological Parameters ◽  
Lung Cancer Patients

Background. MYC (v-myc avian myelocytomatosis viral oncogene homolog) is one of the most frequently amplified genes in lung tumors. For the analysis of gene copy number variations, dPCR (digital PCR) is an appropriate tool. The aim of our study was the assessment of dPCR for the detection of MYC copy number variations (CNV) in lung tissue considering clinicopathological parameters. Material and Methods. MYC status was analyzed with dPCR as well as qPCR (quantitative PCR) using gDNA (genomic DNA) from tumor and adjacent nontumor tissue samples of lung cancer patients. The performance of MYC was estimated based on the AUC (area under curve). Results. The results of the MYC amplification correlated significantly between dPCR and qPCR (rS=0.81, P<0.0001). The MYC copy number revealed by dPCR showed statistically significant differences between tumor and adjacent nontumor tissues. For discrimination, a sensitivity of 43% and a specificity of 99% were calculated, representing 55 true-positive and one false-positive tests. No statistically significant differences could be observed for age, sex, and smoking status or the clinicopathological parameters (histological subtype, grade, and stage). Conclusion. The results of the study show that dPCR is an accurate and reliable method for the determination of MYC copy numbers. The application is characterized by high specificity and moderate sensitivity. MYC amplification is a common event in lung cancer patients, and it is indicated that the determination of the MYC status might be useful in clinical diagnostics.

scholarly journals P3.02b-073 A Phase II, Liquid Biopsy Study Using Digital PCR in EGFR Mutated, Lung Cancer Patients Treated with Afatinib (WJOG 8114LTR)

2017 ◽  
Vol 12 (1) ◽  
pp. S1234 ◽  
Author(s):  
Hiroaki Akamatsu ◽  
Yasuhiro Koh ◽  
Satoshi Morita ◽  
Daichi Fujimoto ◽  
Isamu Okamoto ◽  
...  
Keyword(s):  
Lung Cancer ◽  
Cancer Patients ◽  
Phase Ii ◽  
Liquid Biopsy ◽  
Digital Pcr ◽  
Lung Cancer Patients ◽  
Egfr Mutated

scholarly journals Integrated sequence and gene expression analysis of mouse models of breast cancer reveals critical events with human parallels

2018 ◽  
Author(s):  
Jonathan P Rennhack ◽  
Matthew Swiatnicki ◽  
Yueqi Zhang ◽  
Caralynn Li ◽  
Evan Bylett ◽  
...  
Keyword(s):  
Breast Cancer ◽  
Lung Cancer ◽  
Cancer Patients ◽  
Mouse Models ◽  
Copy Number ◽  
Human Breast Cancer ◽  
Human Cancer ◽  
Copy Number Alterations ◽  
Human Breast ◽  
Lung Cancer Patients

AbstractMouse models have an essential role in cancer research, yet little is known about how various models resemble human cancer at a genomic level. However, the shared genomic alterations in each model and corresponding human cancer are critical for translating findings in mice to the clinic. We have completed whole genome sequencing and transcriptome profiling of two widely used mouse models of breast cancer, MMTV-Neu and MMTV-PyMT. This genomic information was integrated with phenotypic data and CRISPR/Cas9 studies to understand the impact of key events on tumor biology. Despite the engineered initiating transgenic event in these mouse models, they contain similar copy number alterations, single nucleotide variants, and translocation events as human breast cancer. Through integrative in vitro and in vivo studies, we identified copy number alterations in key extracellular matrix proteins including Collagen 1 Type 1 alpha 1 (Col1a1) and Chondroadherin (CHAD) that drive metastasis in these mouse models. Importantly this amplification is also found in 25% of HER2+ human breast cancer and is associated with increased metastasis. In addition to copy number alterations, we observed a propensity of the tumors to modulate tyrosine kinase mediated signaling through mutation of phosphatases. Specifically, we found that 81% of MMTV-PyMT tumors have a mutation in the EGFR regulatory phosphatase, PTPRH. Mutation in PTPRH led to increased phospho-EGFR levels and decreased latency. Moreover, PTPRH mutations increased response to EGFR kinase inhibitors. Analogous PTPRH mutations are present in lung cancer patients and together this data suggests that a previously unidentified population of human lung cancer patients may respond to EGFR targeted therapy. These findings underscore the importance of understanding the complete genomic landscape of a mouse model and illustrate the utility this has in understanding human cancers.

scholarly journals Increased microRNA-30a levels in bronchoalveolar lavage fluid as a diagnostic biomarker for lung cancer

PeerJ ◽  
2021 ◽  
Vol 9 ◽  
pp. e11528
Author(s):  
Wen-Jun Zhu ◽  
Bo-Jiang Chen ◽  
Ying-Ying Zhu ◽  
Ling Sun ◽  
Yu-Chen Zhang ◽  
...  
Keyword(s):  
Lung Cancer ◽  
Bronchoalveolar Lavage ◽  
Cancer Patients ◽  
Cancer Diagnosis ◽  
Roc Curve ◽  
Lung Diseases ◽  
Bronchoalveolar Lavage Fluid ◽  
Lavage Fluid ◽  
Lung Cancer Patients ◽  
Lung Cancer Diagnosis

Background MicroRNA-30a (miRNA-30a) levels have been shown to increase in the plasma of lung cancer patients. Herein, we evaluated the miRNA-30a levels in the bronchoalveolar lavage fluid (BALF) of lung cancer patients as a potential biomarker for lung cancer diagnosis. Methods BALF miRNA-30a expression of 174 subjects was quantified using quantitative real-time reverse transcription-polymerase chain reaction and compared between lung cancer patients and control patients with benign lung diseases. Moreover, its diagnostic value was evaluated by performing receiver operating characteristic (ROC) curve analysis. Results The relative BALF miRNA-30a expression was significantly higher in the lung cancer patients than in the controls (0.74 ±  0.55 versus 0.07 ±  0.48, respectively, p < 0.001) as well as in lung cancer patients with stage I–IIA disease than in those with stage IIB–IV disease (0.98 ±  0.64 versus 0.66 ±  0.54, respectively, p < 0.05). Additionally, miRNA-30a distinguished benign lung diseases from lung cancers, with an area under the ROC curve (AUC) of 0.822. ROC analysis also revealed an AUC of 0.875 for the Youden index-based optimal cut-off points for stage I–IIA adenocarcinoma. Thus, increased miRNA-30a levels in BALF may be a useful biomarker for non-small-cell lung cancer diagnosis.

scholarly journals Case study of an autoantibody panel for early detection of lung cancer and ground-glass nodules

2020 ◽  
Vol 146 (12) ◽  
pp. 3349-3357
Author(s):  
Yunli Huo ◽  
Zijian Guo ◽  
Xuehui Gao ◽  
Zhongjuan Liu ◽  
Ruili Zhang ◽  
...  
Keyword(s):  
Lung Cancer ◽  
Lung Disease ◽  
Cancer Patients ◽  
Cancer Diagnosis ◽  
Predictive Value ◽  
Positive Test ◽  
Test Results ◽  
Newly Diagnosed ◽  
Lung Cancer Patients ◽  
Lung Cancer Diagnosis

Abstract Purpose Increasing lung cancer incidence in China with a high death rate due to late diagnosis highlights the need for biomarkers, such as panels of autoantibodies (AAbs), for prediction and early lung cancer diagnosis. We conducted a study to further evaluate the clinical performance of an AAb diagnostic kit. Methods Using enzyme-linked immunosorbent assay, levels of seven AAbs in serum samples from 121 patients with newly diagnosed lung cancer, 84 controls (34 healthy individuals and 50 patients with benign lung disease), and 100 indeterminate solid nodules, were measured. Participants were followed up until 6 months after a positive test result to confirm lung cancer diagnosis. Results The seven AAb concentration was significantly higher in lung cancer patients than in controls (P < 0.05). The seven AAb sensitivity and specificity for newly diagnosed lung cancer were 45.5% and 85.3%, respectively, while the seven AAb combined area under the curve (in lung cancer patients versus controls) was 0.660. Of the 28 patients with solid nodules with positive test results, 8 and 3 were diagnosed with lung cancer and benign lung disease, respectively, during follow-up. The positive predictive value of the experiment was 72.7%. Conclusion Positive AAb test results were associated with a high risk of lung cancer. The seven-AAb panel also had a high predictive value for detecting lung cancer in patients with solid nodules. Our seven lung cancer autoantibody types can provide an important early warning sign in the clinical setting.

scholarly journals A CRISPR Test for Rapidly and Sensitively Detecting Circulating EGFR Mutations

Diagnostics ◽  
2020 ◽  
Vol 10 (2) ◽  
pp. 114 ◽  
Author(s):  
Jen-Hui Tsou ◽  
Qixin Leng ◽  
Feng Jiang
Keyword(s):  
Lung Cancer ◽  
Cancer Patients ◽  
Limit Of Detection ◽  
Digital Pcr ◽  
Medical Diagnostics ◽  
Egfr Mutations ◽  
Nucleic Acid Detection ◽  
Great Promise ◽  
Plasma Samples ◽  
Lung Cancer Patients

The detection of EGFR mutations in circulating cell-free DNA can enable personalized therapy for cancer. The current techniques for detecting circulating EGFR mutations are expensive and time-consuming with moderate sensitivity. Emerging CRISPR is revolutionizing medical diagnostics and showing a great promise for nucleic acid detection. This study aims to develop CRISPR-Cas12a as a simple test to sensitively detect circulating EGFR mutations in plasma. Serially diluted samples of DNA containing heterozygous EGFR mutations (L858R and T790M) in wild-type genomic DNA are concurrently tested for the mutations by a CRISPR-Cas12a system and droplet digital PCR (ddPCR). The CRISPR-Cas12a system can detect both L858R and T790M with a limit of detection of 0.005% in less than three hours. ddPCR detects the mutations with a limit of detection of 0.05% for more than five hours. Plasma samples of 28 lung cancer patients and 20 cancer-free individuals are tested for the EGFR mutations by CRISPR-Cas12a system and ddPCR. The CRISPR-Cas12a system could detect L858R in plasma of two lung cancer patients whose tissue biopsies are positive for L858R, and one plasma sample of three lung cancer patients whose tissue biopsies are positive for T790M. ddPCR detects L858R in the same two plasm samples, however, does not detect T790M in any of the plasma samples. This proof of principle study demonstrates that the CRISPR-Cas12a system could rapidly and sensitively detect circulating EGFR mutations, and thus, has potential prognostic or therapeutic implications.

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