Quantification of messenger ribonucleic acid for epidermal growth factor in human myometrium and leiomyomata using reverse transcriptase polymerase chain reaction

1994 ◽  
Vol 78 (5) ◽  
pp. 1179-1184 ◽  
Author(s):  
M. L. Harrison-Woolrych
Keyword(s):  
Polymerase Chain Reaction ◽  
Growth Factor ◽  
Epidermal Growth Factor ◽  
Reverse Transcriptase ◽  
Ribonucleic Acid ◽  
Human Myometrium ◽  
Chain Reaction ◽  
Messenger Ribonucleic Acid ◽  
Epidermal Growth ◽  
Polymerase Chain

scholarly journals Development of real-time reverse transcription polymerase chain reaction assays to quantify insulin-like growth factor receptor and insulin receptor expression in equine tissue

2013 ◽  
Vol 80 (1) ◽  
Author(s):  
Stephen B. Hughes ◽  
Melvyn Quan ◽  
Alan Guthrie ◽  
Martin Schulman
Keyword(s):  
Polymerase Chain Reaction ◽  
Growth Factor ◽  
Insulin Receptor ◽  
Ribonucleic Acid ◽  
Muscle Metabolism ◽  
Insulin Like Growth Factor ◽  
Chain Reaction ◽  
Messenger Ribonucleic Acid ◽  
Receptor Assay ◽  
Polymerase Chain

The insulin-like growth factor system (insulin-like growth factor 1, insulin-like growth factor 2, insulin-like growth factor 1 receptor, insulin-like growth factor 2 receptor and six insulin-like growth factor-binding proteins) and insulin are essential to muscle metabolism and most aspects of male and female reproduction. Insulin-like growth factor and insulin play important roles in the regulation of cell growth, differentiation and the maintenance of cell differentiation in mammals. In order to better understand the local factors that regulate equine physiology, such as muscle metabolism and reproduction (e.g., germ cell development and fertilisation), real-time reverse transcription polymerase chain reaction assays for quantification of equine insulin-like growth factor 1 receptor and insulin receptor messenger ribonucleic acid were developed. The assays were sensitive: 192 copies/µLand 891 copies/µL for insulin-like growth factor 1 receptor, messenger ribonucleic acid and insulin receptor respectively (95%limit of detection), and efficient: 1.01 for the insulin-like growth factor 1 receptor assay and 0.95 for the insulin receptor assay. The assays had a broad linear range of detection (seven logs for insulin-like growth factor 1 receptor and six logs for insulin receptor). This allowed for analysis of very small amounts of messenger ribonucleic acid. Low concentrations of both insulin-like growth factor 1 receptor and insulin receptor messenger ribonucleic acid were detected in endometrium, lung and spleen samples, whilst high concentrations were detected in heart, muscle and kidney samples, this was most likely due to the high level of glucose metabolism and glucose utilisation by these tissues. The assays developed for insulin-like growth factor 1 receptor and insulin receptor messenger ribonucleic acid expression have been shown to work on equine tissue and will contribute to the understanding of insulin and insulin-like growth factor 1 receptor physiology in the horse.

scholarly journals Exosomes as a biomarker platform for detecting epidermal growth factor receptor–positive high-grade gliomas

2018 ◽  
Vol 128 (4) ◽  
pp. 1091-1101 ◽  
Author(s):  
Sasidhar Venkata Manda ◽  
Yogesh Kataria ◽  
Babul Reddy Tatireddy ◽  
Balasubramaniam Ramakrishnan ◽  
Boola Gnana Ratnam ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Epidermal Growth Factor Receptor ◽  
Growth Factor ◽  
Epidermal Growth Factor ◽  
Growth Factor Receptor ◽  
High Grade ◽  
Chain Reaction ◽  
Epidermal Growth ◽  
Polymerase Chain ◽  
Paired Serum

OBJECTIVEHigh-grade glial brain tumors are often characterized by an elevated expression of the tumorigenic epidermal growth factor receptor variant III (EGFRvIII). The authors sought to establish a clinically adaptive protocol as a noninvasive diagnostic tool for EGFRvIII detection through serum exosomes.METHODSPurity of serum exosome/RNA was confirmed by electron microscopy and flow cytometry and through an RNA bioanalyzer profile. EGFRvIII amplification was initially established by semiquantitative polymerase chain reaction in tumor tissues and exosomes. Diagnostic performance of EGFRvIII transcript in tissue versus exosome was determined using a 2 × 2 clinical table approach. Overall survival was determined using Kaplan-Meier analysis.RESULTSThe EGFRvIII transcript was detected in 39.5% of tumor tissue samples and in 44.7% of their paired serum exosome samples; 28.1% of biopsy tumors coexpressed wild-type EGFR and EGFRvIII. Tissue EGFRvIII amplification served as the reference-positive control for its paired serum expression. The overall clinical sensitivity and specificity of semiquantitative exosome EGFRvIII polymerase chain reaction detection assay in serum were 81.58% (95% CI 65.67%–92.26%) and 79.31% (95% CI 66.65%–88.83%), respectively. Age, sex, tumor location, and side of the body on which the tumor was located had no effect on the detection rate of exosomal EGFRvIII transcript. EGFRvIII expression either in exosomes or tissue correlated with poor survival.CONCLUSIONSThe authors established a serum-based method for detection of EGFRvIII in high-grade brain tumors that might serve as an optimal noninvasive method for diagnosing EGFRvIII-positive high-grade gliomas.

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