scholarly journals Locked nucleic acid inhibits amplification of contaminating DNA in real-time PCR

BioTechniques ◽  
2005 ◽  
Vol 38 (4) ◽  
pp. 605-610 ◽  
Author(s):  
Lone Hummelshoj ◽  
Lars P. Ryder ◽  
Hans O. Madsen ◽  
Lars K. Poulsen
Keyword(s):  
Nucleic Acid ◽  
Real Time ◽  
Real Time Pcr ◽  
Locked Nucleic Acid

scholarly journals Locked Nucleic Acid Probe-Based Real-Time PCR Assay for the Rapid Detection of Rifampin-Resistant Mycobacterium tuberculosis

PLoS ONE ◽  
2015 ◽  
Vol 10 (11) ◽  
pp. e0143444 ◽  
Author(s):  
Yong Zhao ◽  
Guilian Li ◽  
Chongyun Sun ◽  
Chao Li ◽  
Xiaochen Wang ◽  
...  
Keyword(s):  
Mycobacterium Tuberculosis ◽  
Nucleic Acid ◽  
Real Time ◽  
Real Time Pcr ◽  
Rapid Detection ◽  
Locked Nucleic Acid ◽  
Pcr Assay ◽  
Locked Nucleic Acid Probe

scholarly journals Detection of Spinal Muscular Atrophy Using a Duplexed Real-Time PCR Approach With Locked Nucleic Acid-Modified Primers

2021 ◽  
Vol 41 (1) ◽  
pp. 101-107
Author(s):  
Jianyan Pan ◽  
Chunhua Zhang ◽  
Yanling Teng ◽  
Sijing Zeng ◽  
Siyi Chen ◽  
...  
Keyword(s):  
Nucleic Acid ◽  
Spinal Muscular Atrophy ◽  
Real Time ◽  
Real Time Pcr ◽  
Muscular Atrophy ◽  
Locked Nucleic Acid

scholarly journals Use of locked nucleic acid oligonucleotides as hybridization/FRET probes for quantification of 16S rDNA by real-time PCR

BioTechniques ◽  
2005 ◽  
Vol 38 (1) ◽  
pp. 29-31 ◽  
Author(s):  
Oliver Goldenberg ◽  
Olfert Landt ◽  
Ralf R. Schumann ◽  
Ulf B. Göbel ◽  
Lutz Hamann
Keyword(s):  
Nucleic Acid ◽  
Real Time ◽  
16S Rdna ◽  
Real Time Pcr ◽  
Locked Nucleic Acid

Fast and sensitive real-time PCR detection of major antiviral drug-resistance mutations in chronic hepatitis B patients using a predesigned panel of Locked-Nucleic-Acid TaqMan probes

2021 ◽  
Author(s):  
Son V. Chu ◽  
Son T. Vu ◽  
Hang M. Nguyen ◽  
Ngan T. Le ◽  
Phuong T. Truong ◽  
...  
Keyword(s):  
Drug Resistance ◽  
Chronic Hepatitis ◽  
Nucleic Acid ◽  
Hepatitis B ◽  
Real Time ◽  
Chronic Hepatitis B ◽  
Real Time Pcr ◽  
Locked Nucleic Acid ◽  
Resistance Mutations ◽  
Drug Resistance Mutations

We developed a novel real-time PCR assay that simultaneously evaluates eleven major nucleos(t)ide antiviral (NA) drug-resistance mutations (mt) in chronic hepatitis B patients (CHB), including L180M, M204I/V, and V207M (lamivudine (LMV) resistance), N/H238A/T (Adefovir (ADF) resistance) which are circulating in Vietnam; T184G/L, S202I, M250V (entecavir (ETV) resistance), and A194T (tenofovir resistance) which have been recently reported in several studies across the globe. We detected drug-resistant mt in HBV samples using our predesigned panel of allele-specific locked-nucleic acid (LNA) probes. Our assay had a high sensitivity of 5% in a low HBV DNA population of ≥5 ×10 3 IU/mL and was validated in a cohort of 130 treatment-naive children and 98 NA-experienced adults with CHB. Single-point mt for LMV- and ADF-resistance were detected in 57.7% and 54.1% of the child and adult samples, respectively, with rtV207M (42.3%, children; 36.7%, adults) and rtN238T/A (15.4%, children; 16.3, adults) being the most frequent mt in these populations. Multiple-point mt including rtL180M-rtM204V- rtN238A and rtL180M-rtM204I were only identified in two children, resulting in LMV-ADF-resistance and reduced ETV susceptibility. In conclusion, this assay accurately identified the mt profile of children (98.4%) and adults (91.2%) with CHB, which is comparable to established methods. This fast and sensitive screening method can be used for the detection of major NA-resistant mt circulating in developing countries, as well as providing a model for the development of similar mt-detection assays, especially for use in non-hospitalised patients who need their results within half a day before starting treatment.

Multiplex TaqMan locked nucleic acid real-time PCR for the differential identification of various meat and meat products

Meat Science ◽  
2018 ◽  
Vol 137 ◽  
pp. 41-46 ◽  
Author(s):  
Rusu Xu ◽  
Shuang Wei ◽  
Guangbiao Zhou ◽  
Jiao Ren ◽  
Zhongyong Liu ◽  
...  
Keyword(s):  
Nucleic Acid ◽  
Real Time ◽  
Real Time Pcr ◽  
Meat Products ◽  
Locked Nucleic Acid ◽  
Differential Identification

scholarly journals Library of Prefabricated Locked Nucleic Acid Hydrolysis Probes Facilitates Rapid Development of Reverse-Transcription Quantitative Real-Time PCR Assays for Detection of Novel Influenza A/H1N1/09 Virus

2009 ◽  
Vol 55 (12) ◽  
pp. 2218-2222 ◽  
Author(s):  
Jürgen J Wenzel ◽  
Heiko Walch ◽  
Markus Bollwein ◽  
Hans Helmut Niller ◽  
Waltraud Ankenbauer ◽  
...  
Keyword(s):  
Nucleic Acid ◽  
Real Time ◽  
Reverse Transcription ◽  
Real Time Pcr ◽  
Influenza A ◽  
Rapid Development ◽  
Locked Nucleic Acid ◽  
Quantitative Real Time Pcr ◽  
Influenza A H1n1 ◽  
Novel Influenza A

Abstract Background: The emergence of a novel pandemic human strain of influenza A (H1N1/09) has clearly demonstrated the need for flexible tools enabling the rapid development of new diagnostic methods. Methods: We designed a set of reverse-transcription quantitative real-time PCR (RT-qPCR) assays based on the Universal ProbeLibrary (UPL)—a collection of 165 presynthesized, fluorescence-labeled locked nucleic acid (LNA) hydrolysis probes—specifically to detect the novel influenza A virus. We evaluated candidate primer/UPL-probe pairs with 28 novel influenza A/H1N1/09 patient samples of European and Mexican origin. Results: Of 14 assays in the hemagglutinin (HA) and neuraminidase (NA) genes, 12 detected viral nucleic acids from diluted patient samples without need for further optimization. We characterized the diagnostic specificity of the 2 best-performing assays with a set of samples comprising various influenza virus strains of human and animal origin that showed no cross-reactivity. The diagnostic sensitivity of these 2 primer/probe combinations was in the range of 100–1000 genomic copies/mL. In comparison to a reference assay recommended by the German health authorities, the analytical sensitivities and specificities of the assays were equivalent. Conclusions: Facing the emergence of novel influenza A/H1N1/09, we were able to develop, within 2 days, a set of sensitive and specific RT-qPCR assays for the laboratory diagnosis of suspected cases. H1N1/09 served as a model to show the feasibility of the UPL approach for the expedited development of new diagnostic assays to detect emerging pathogens.

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