Identification of squid species by melting temperature shifts on fluorescence melting curve analysis (FMCA) using single dual-labeled probe

2017 ◽  
Author(s):  
Eunjung Koh ◽  
Ha Jeong Song ◽  
Na Young Kwon ◽  
Gi Won Kim ◽  
Kwang Ho Lee ◽  
...  
Keyword(s):  
Melting Temperature ◽  
Melting Curve ◽  
Curve Analysis ◽  
Melting Curve Analysis ◽  
Squid Species ◽  
Fluorescence Melting Curve Analysis ◽  
Labeled Probe

Probe-Based Fluorescence Melting Curve Analysis for Differentiating Larimichthys polyactis and Larimichthys crocea

2016 ◽  
Vol 9 (7) ◽  
pp. 2036-2041 ◽  
Author(s):  
Jeong Jin Ahn ◽  
Youngjoo Kim ◽  
Ji Young Hong ◽  
Gi Won Kim ◽  
Seol Young Kim ◽  
...  
Keyword(s):  
Melting Curve ◽  
Curve Analysis ◽  
Melting Curve Analysis ◽  
Larimichthys Crocea ◽  
Larimichthys Polyactis ◽  
Fluorescence Melting Curve Analysis

scholarly journals Detection of Eight Respiratory Bacterial Pathogens Based on Multiplex Real-Time PCR with Fluorescence Melting Curve Analysis

2020 ◽  
Vol 2020 ◽  
pp. 1-11
Author(s):  
Liuyang Hu ◽  
Bing Han ◽  
Qin Tong ◽  
Hui xiao ◽  
Donglin Cao
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Limit Of Detection ◽  
Bacterial Pathogens ◽  
Melting Curve ◽  
Cost Effective ◽  
Culture Method ◽  
Curve Analysis ◽  
Melting Curve Analysis ◽  
Fluorescence Melting Curve Analysis

Background and Objective. Acinetobacter baumannii, Escherichia coli, Klebsiella pneumoniae, Streptococcus pneumoniae, Haemophilus influenzae, Staphylococcus aureus, Pseudomonas aeruginosa, and Mycobacterium tuberculosis are primary respiratory bacterial pathogens contributing to morbidity and mortality in developing countries. This study evaluated the diagnostic performance of multiplex real-time PCR with fluorescence melting curve analysis (MCA) assay, which was used to detect eight respiratory bacterial pathogens simultaneously. Methods. A total of 157 sputum specimens were examined by multiplex real-time with fluorescence MCA, and the results were compared with the conventional culture method. Results. Multiplex real-time PCR with fluorescence MCA specifically detected and differentiated eight respiratory bacterial pathogens by different melting curve peaks for each amplification product within 2 hours and exhibited high repeatability. The limit of detection ranged from 64 to 102 CFU/mL in the multiplex PCR system. Multiplex real-time PCR with fluorescence MCA showed a sensitivity greater than 80% and a 100% specificity for each pathogen. The kappa correlation of eight bacteria ranged from 0.89 to 1.00, and the coefficient of variation ranged from 0.05% to 0.80%. Conclusions. Multiplex real-time PCR with fluorescence MCA assay is a sensitive, specific, high-throughput, and cost-effective method to detect multiple bacterial pathogens simultaneously.

scholarly journals Web-based LinRegPCR: application for the visualization and analysis of (RT)-qPCR amplification and melting data

2021 ◽  
Vol 22 (1) ◽  
Author(s):  
Andreas Untergasser ◽  
Jan M. Ruijter ◽  
Vladimir Benes ◽  
Maurice J. B. van den Hoff
Keyword(s):  
Melting Temperature ◽  
Melting Curve ◽  
Curve Analysis ◽  
Melting Curve Analysis ◽  
Amplification Product ◽  
Web Based ◽  
Amplification Curve ◽  
Qpcr Data ◽  
Melting Curves ◽  
The Individual

Abstract Background The analyses of amplification and melting curves have been shown to provide valuable information on the quality of the individual reactions in quantitative PCR (qPCR) experiments and to result in more reliable and reproducible quantitative results. Implementation The main steps in the amplification curve analysis are (1) a unique baseline subtraction, not using the ground phase cycles, (2) PCR efficiency determination from the exponential phase of the individual reactions, (3) setting a common quantification threshold and (4) calculation of the efficiency-corrected target quantity with the common threshold, efficiency per assay and Cq per reaction. The melting curve analysis encompasses smoothing of the observed fluorescence data, normalization to remove product-independent fluorescence loss, peak calling and assessment of the correct peak by comparing its melting temperature with the known melting temperature of the intended amplification product. Results The LinRegPCR web application provides visualization and analysis of a single qPCR run. The user interface displays the analysis results on the amplification curve analysis and melting curve analysis in tables and graphs in which deviant reactions are highlighted. The annotated results in the tables can be exported for calculation of gene-expression ratios, fold-change between experimental conditions and further statistical analysis. Web-based LinRegPCR addresses two types of users, wet-lab scientists analyzing the amplification and melting curves of their own qPCR experiments and bioinformaticians creating pipelines for analysis of series of qPCR experiments by splitting its functionality into a stand-alone back-end RDML (Real-time PCR Data Markup Language) Python library and several companion applications for data visualization, analysis and interactive access. The use of the RDML data standard enables machine independent storage and exchange of qPCR data and the RDML-Tools assist with the import of qPCR data from the files exported by the qPCR instrument. Conclusions The combined implementation of these analyses in the newly developed web-based LinRegPCR (https://www.gear-genomics.com/rdml-tools/) is platform independent and much faster than the original Windows-based versions of the LinRegPCR program. Moreover, web-based LinRegPCR includes a novel statistical outlier detection and the combination of amplification and melting curve analyses allows direct validation of the amplification product and reporting of reactions that amplify artefacts.

A new method for ABO genotyping using fluorescence melting curve analysis based on peptide nucleic acid probes

2015 ◽  
Vol 17 (5) ◽  
pp. 334-339 ◽  
Author(s):  
Kyungmyung Lee ◽  
Hyun-Chul Park ◽  
Sanghyun An ◽  
Eu-Ree Ahn ◽  
Yang-Han Lee ◽  
...  
Keyword(s):  
Nucleic Acid ◽  
Peptide Nucleic Acid ◽  
Melting Curve ◽  
Curve Analysis ◽  
New Method ◽  
Melting Curve Analysis ◽  
Nucleic Acid Probes ◽  
Fluorescence Melting Curve Analysis

Detection of vaccinia virus DNA on the LightCycler by fluorescence melting curve analysis

2005 ◽  
Vol 126 (1-2) ◽  
pp. 187-195 ◽  
Author(s):  
Andreas Nitsche ◽  
Brigitte Steger ◽  
Heinz Ellerbrok ◽  
Georg Pauli
Keyword(s):  
Vaccinia Virus ◽  
Melting Curve ◽  
Curve Analysis ◽  
Melting Curve Analysis ◽  
Fluorescence Melting Curve Analysis
Sign in / Sign up

Export Citation Format

Share Document