scholarly journals Endonuclease Restriction-Mediated Real-Time Polymerase Chain Reaction: A Novel Technique for Rapid, Sensitive and Quantitative Detection of Nucleic-Acid Sequence

2016 ◽  
Vol 7 ◽  
Author(s):  
Yi Wang ◽  
Yan Wang ◽  
Lu Zhang ◽  
Machao Li ◽  
Lijuan Luo ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Nucleic Acid ◽  
Real Time ◽  
Nucleic Acid Sequence ◽  
Quantitative Detection ◽  
Chain Reaction ◽  
Novel Technique ◽  
Polymerase Chain

Quantitative Real-Time Polymerase Chain Reaction Detection of BK Virus Using Labeled Primers

2010 ◽  
Vol 134 (3) ◽  
pp. 444-448 ◽  
Author(s):  
Zhengming Gu ◽  
Jianmin Pan ◽  
Matthew J. Bankowski ◽  
Randall T. Hayden
Keyword(s):  
Polymerase Chain Reaction ◽  
Real Time ◽  
Real Time Pcr ◽  
Quantitative Polymerase Chain Reaction ◽  
Bk Virus ◽  
Clinical Samples ◽  
Quantitative Detection ◽  
Chain Reaction ◽  
Pcr Method ◽  
Polymerase Chain

Abstract Context.—BK virus infections among immunocompromised patients are associated with disease of the kidney or urinary bladder. High viral loads, determined by quantitative polymerase chain reaction (PCR), have been correlated with clinical disease. Objective.—To develop and evaluate a novel method for real-time PCR detection and quantification of BK virus using labeled primers. Design.—Patient specimens (n = 54) included 17 plasma, 12 whole blood, and 25 urine samples. DNA was extracted using the MagNA Pure LC Total Nucleic Acid Isolation Kit (Roche Applied Science, Indianapolis, Indiana); sample eluate was PCR-amplified using the labeled primer PCR method. Results were compared with those of a user-developed quantitative real-time PCR method (fluorescence resonance energy transfer probe hybridization). Results.—Labeled primer PCR detected less than 10 copies per reaction and showed quantitative linearity from 101 to 107 copies per reaction. Analytical specificity of labeled primer PCR was 100%. With clinical samples, labeled primer PCR demonstrated a trend toward improved sensitivity compared with the reference method. Quantitative assay comparison showed an R2 value of 0.96 between the 2 assays. Conclusions.—Real-time PCR using labeled primers is highly sensitive and specific for the quantitative detection of BK virus from a variety of clinical specimens. These data demonstrate the applicability of labeled primer PCR for quantitative viral detection and offer a simplified method that removes the need for separate oligonucleotide probes.

Quantitative detection of HCMV-DNA in saliva from infants and breast milk on real-time polymerase chain reaction

2009 ◽  
Vol 51 (4) ◽  
pp. 530-534 ◽  
Author(s):  
Hiroaki Murata ◽  
Ritsue Nii ◽  
Masahiro Ito ◽  
Toshiaki Ihara ◽  
Yoshihiro Komada
Keyword(s):  
Polymerase Chain Reaction ◽  
Breast Milk ◽  
Real Time ◽  
Quantitative Detection ◽  
Chain Reaction ◽  
Polymerase Chain

Ribonuclease H-cleavable and recombinase-quenching fluorescent probes for the real-time detection of strand invasion based amplification

2019 ◽  
Vol 11 (43) ◽  
pp. 5568-5576
Author(s):  
Sonja Elf ◽  
Kevin E. Eboigbodin
Keyword(s):  
Polymerase Chain Reaction ◽  
Nucleic Acid ◽  
Real Time ◽  
Fluorescent Probes ◽  
Nucleic Acid Amplification ◽  
Ribonuclease H ◽  
Chain Reaction ◽  
Amplification Method ◽  
Polymerase Chain ◽  
Strand Invasion

SIBA is an established nucleic acid amplification method that is used as an alternative to polymerase chain reaction (PCR).

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