scholarly journals A New Approach for Accurate Identification of Allele State Based on Real-Time PCR for Biomedical Tasks

2022 ◽  
Author(s):  
Ernest Benaguev ◽  
Ivan Vladimirov ◽  
Olga Pavlova ◽  
Denis Bogomaz
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Diagnostic System ◽  
Significant Snps ◽  
Nucleotide Polymorphisms ◽  
Accurate Identification ◽  
New Approach ◽  
Conventional Systems ◽  
Fine Tune

Genotyping of single nucleotide polymorphisms (SNPs) is an important task in medicine, veterinary medicine and biology. Precise differentiation of SNPs can be challenging. Methods based onTaqman can lead to false positive results due to nonspecific annealing of the probe. The aim of this research was to develop a new approach for the accurate differentiation of SNPs based on real-time PCR with Taqmanprobes and their rivals.The rivals competed with the Taqmanprobes for annealing to the site. The rivals blocked the nonspecific allele so that the Taqmanprobe could not anneal to it. Thus,the Taqmanprobe only detected specific alleles.This approach madeit possible to fine-tune the diagnostic system by selecting the ratio of Taqmanprobes and rivals (in non-equimolar amounts too).The new approach was tested on several diagonally significant SNPs in veterinary medicine.Using Taqman probes and rival probes showed a significantly greater specificity and efficiency in the determination of both homozygotes and heterozygotes than when conventional systems based only on Taqmanwere used. Keywords: SNP, allele identification, real-time PCR, fluorescent dye

Real-Time PCR Determination of Escherichia coli Genomic DNA Contamination in Plasmid Preparations

2002 ◽  
Vol 301 (1) ◽  
pp. 151-153 ◽  
Author(s):  
Adrián Vilalta ◽  
Vanessa Whitlow ◽  
Terrie Martin
Keyword(s):  
Escherichia Coli ◽  
Real Time ◽  
Real Time Pcr ◽  
Genomic Dna ◽  
Dna Contamination

Multiplex real-time PCR for the detection of insect DNA and determination of contents of Tenebrio molitor, Locusta migratoria and Achaeta domestica in food

2019 ◽  
Vol 245 (3) ◽  
pp. 559-567 ◽  
Author(s):  
René Köppel ◽  
Rafael Schum ◽  
Michael Habermacher ◽  
Cindy Sester ◽  
Lucia Eugeni Piller ◽  
...  
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Locusta Migratoria ◽  
Tenebrio Molitor

scholarly journals Real-Time PCR as a Versatile Tool for Investigating the Susceptibility of Human Herpesvirus 6 to Antiviral Agents

2003 ◽  
Vol 47 (9) ◽  
pp. 3021-3024 ◽  
Author(s):  
Muriel Macé ◽  
Chaysavanh Manichanh ◽  
Pascale Bonnafous ◽  
Stéphanie Précigout ◽  
David Boutolleau ◽  
...  
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Antiviral Agents ◽  
Antiviral Drug ◽  
Human Herpesvirus ◽  
Drug Susceptibility ◽  
Human Herpesvirus 6 ◽  
Versatile Tool ◽  
Herpesvirus 6

ABSTRACT A quantitative real-time PCR assay was developed for the determination of antiviral drug susceptibility and growth kinetics of human herpesvirus 6. The susceptibility and fitness of a sensitive strain, HST, and its ganciclovir-resistant derivative, GCVR1, were then characterized, leading us to conclude that the mutations of this latter virus did not alter its fitness significantly.

scholarly journals Duplex Real-Time PCR Assay for Rapid Detection of Ampicillin-Resistant Enterococcus faecium

2004 ◽  
Vol 48 (2) ◽  
pp. 556-560 ◽  
Author(s):  
Stein Christian Mohn ◽  
Arve Ulvik ◽  
Roland Jureen ◽  
Rob J. L. Willems ◽  
Janetta Top ◽  
...  
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Enterococcus Faecium ◽  
Rapid Detection ◽  
Resistant Bacteria ◽  
Pcr Assay ◽  
Culture Methods ◽  
Accurate Identification ◽  
Antibiotic Resistant ◽  
Highly Correlated

ABSTRACT Rapid and accurate identification of carriers of resistant microorganisms is an important aspect of efficient infection control in hospitals. Traditional identification methods of antibiotic-resistant bacteria usually take at least 3 to 4 days after sampling. A duplex real-time PCR assay was developed for rapid detection of ampicillin-resistant Enterococcus faecium (ARE). Primers and probes that are used in this assay specifically detected the d-Ala-d-Ala ligase gene of E. faecium and the modified penicillin-binding protein 5 gene (pbp5) carrying the Glu-to-Val substitution at position 629 (Val-629) in a set of 129 tested E. faecium strains with known pbp5 sequence. Presence of the Val-629 in the strain set from 11 different countries was highly correlated with ampicillin resistance. In a screening of hospitalized patients, the real-time PCR assay yielded a sensitivity and a specificity for the detection of ARE colonization of 95% and 100%, respectively. The results were obtained 4 h after samples were harvested from overnight broth of rectal swab samples, identifying both species and the resistance marker mutation in pbp5. This novel assay reliably identifies ARE 2 to 3 days more quickly than traditional culture methods, thereby increasing laboratory throughput, making it useful for rectal screening of ARE. The assay demonstrates the advantages of real-time PCR for detection of nosocomial pathogens.

Determination of Helicobacter pylori cagA, vacA genotypes with real-time PCR melting curve analysis

2001 ◽  
Vol 95 (1-6) ◽  
pp. 369-377 ◽  
Author(s):  
Agnes Ruzsovics ◽  
Bela Molnar ◽  
Zsuzsa Unger ◽  
Zsolt Tulassay ◽  
Laszlo Pronai
Keyword(s):  
Helicobacter Pylori ◽  
Real Time ◽  
Real Time Pcr ◽  
Melting Curve ◽  
Curve Analysis ◽  
Melting Curve Analysis

scholarly journals A one-step method for quantitative determination of uracil in DNA by real-time PCR

2010 ◽  
Vol 38 (21) ◽  
pp. e196-e196 ◽  
Author(s):  
András Horváth ◽  
Beáta G. Vértessy
Keyword(s):  
Quantitative Determination ◽  
Real Time ◽  
Real Time Pcr ◽  
Step Method ◽  
One Step
Sign in / Sign up

Export Citation Format

Share Document