Rapid and inexpensive species differentiation using a multiplex real-time polymerase chain reaction high-resolution melt assay

2016 ◽  
Vol 500 ◽  
pp. 15-17 ◽  
Author(s):  
Kelly M. Elkins ◽  
Anjelica C.U. Perez ◽  
Katherine C. Sweetin
Keyword(s):  
Polymerase Chain Reaction ◽  
High Resolution ◽  
Real Time ◽  
Species Differentiation ◽  
Chain Reaction ◽  
High Resolution Melt ◽  
Polymerase Chain

scholarly journals Comparison of Direct Sequencing with Real-time PCR High Resolution Melt and PCR Restriction Fragment Length Polymorphism Analysis to Identify Clinically Important Candida Species

2021 ◽  
Vol 16 (4) ◽  
Author(s):  
Zeynab Yassin ◽  
Fariba Shirvani ◽  
Mahsa Fattahi
Keyword(s):  
Polymerase Chain Reaction ◽  
High Resolution ◽  
Real Time ◽  
Fragment Length ◽  
Sanger Sequencing ◽  
Chain Reaction ◽  
High Resolution Melt ◽  
Pcr Rflp ◽  
Polymerase Chain ◽  
Restriction Fragment Length

Background: Candida albicans is the predominant yeast reported from human infection. Non-albicans Candida species have been recently developed as medically vital fungi. Therefore, it is essential to detect and identify the pathogens at the species level to prescribe appropriate treatment. Methods: This study assessed two complementary methods, including real-time polymerase chain reaction-high resolution melt (PCR-HRM) and polymerase chain reaction-restriction fragment length morphism (PCR-RFLP) with standard PCR and Sanger sequencing as the benchmark. Results: In total, 66 samples were tested, and two newly-advanced assays were more effective and displayed comprehensive concordance (66/66, 100%) with Sanger sequencing outcomes. Moreover, accurate and economical tests were positively advanced by real-time PCR-HRM for C. albicans and C. parapsilosis complexes. Conclusions: Given the number of studies performed on the comparison of sensitivity and specificity of phenotypic and genotypic methods to diagnose and identify invasive fungal pathogens and the findings of this study, it could be stated that the correlative PCR-HRM and PCR-RFLP methods were effectively advanced as substitutes for conventional Sanger sequencing for the reasonable identification. However, supplementary evaluations and confirming studies should be carried out with a broad range of samples to standardize this method for routine application in medical laboratories.

Real-time PCR and high-resolution melt analysis methods for detection of pathogenic species of Brucella

2017 ◽  
Vol 41 (6) ◽  
Author(s):  
Faramarz Masjedian Jazi ◽  
Reza Mirnejad ◽  
Vahhab Piranfar ◽  
Noor Amir Mozafari ◽  
Taghi Zahraei Salehi ◽  
...  
Keyword(s):  
High Resolution ◽  
Real Time ◽  
Serum Antibody ◽  
Clinical Samples ◽  
Species Differentiation ◽  
High Resolution Melt ◽  
High Resolution Melt Analysis ◽  
Antibody Titers ◽  
Polymerase Chain ◽  
Animal Isolates

AbstractBackground:It is of great importance to quickly and accurately detectMethods:The current study describes a new method for the detection of brucellosis in clinical samples using real-time polymerase chain reaction (PCR) and high-resolution melt (HRM) curve analysis. This study was conducted on 70 human and 55 animal isolates with more than 1/80 serum antibody titers. Additionally, the accuracy and specificity of the methods were compared.Results:The mean range [cycles threshold±standard deviation (CConclusions:The results of HRM analysis can be used for species differentiation and bacterial genotyping according to nucleotide polymorphism. This molecular method could help in diagnosing

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