Smartphone-based chemiluminometric hybridization assays and quantitative competitive polymerase chain reaction

2019 ◽  
Vol 1088 ◽  
pp. 123-130 ◽  
Author(s):  
Panagiota M. Kalligosfyri ◽  
Areti Sevastou ◽  
Iraklis K. Kyriakou ◽  
Sotirios S. Tragoulias ◽  
Despina P. Kalogianni ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Chain Reaction ◽  
Competitive Polymerase Chain Reaction ◽  
Polymerase Chain ◽  
Hybridization Assays

Differential display competitive polymerase chain reaction: An optimal tool for assaying gene expression

1999 ◽  
Vol 20 (2) ◽  
pp. 230 ◽  
Author(s):  
Marianne Jorgensen ◽  
Maja Bévort ◽  
Thuri S. Kledal ◽  
Brian V. Hansen ◽  
Marlene Dalgaard ◽  
...  
Keyword(s):  
Gene Expression ◽  
Polymerase Chain Reaction ◽  
Differential Display ◽  
Chain Reaction ◽  
Competitive Polymerase Chain Reaction ◽  
Polymerase Chain

scholarly journals Detection and Differentiation of Bovine Group A Rotavirus Serotypes using Polymerase Chain Reaction-Generated Probes to the VP7 Gene

1992 ◽  
Vol 4 (2) ◽  
pp. 148-158 ◽  
Author(s):  
Anil V. Parwani ◽  
Blair I. Rosen ◽  
Jorge Flores ◽  
Malcolm A. McCrae ◽  
Mario Gorziglia ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Blot Hybridization ◽  
Northern Blot Hybridization ◽  
Chain Reaction ◽  
Diarrhea Virus ◽  
Bovine Rotavirus ◽  
Group A ◽  
Polymerase Chain ◽  
Vp7 Gene ◽  
Hybridization Assays

Dot and Northern blot hybridization assays were developed to detect and differentiate group A bovine rotavirus serotypes using radiolabeled serotype 6 (Nebraska calf diarrhea virus [NCDV] and United Kingdom [UK] strains) or serotype 10 (Cracker [Cr] strain) VP7 gene probes. Partial length VP7-specific cDNA encompassing areas of major sequence diversity were generated by the polymerase chain reaction (PCR) using either cloned VP7 genes (NCDV and UK strains) or reverse transcribed mRNA (Cr strain) as templates. Radiolabeled probes prepared from the PCR-generated cDNA were tested at various stringency conditions to optimize the hybridization assays. At high stringency conditions (52 C, 50% formamide, 5 x standard saline citrate), the NCDV, UK, and Cr probes serotypically differentiated bovine rotavirus isolates in RNA samples prepared from cell culture-propagated viruses or in fecal specimens from infected gnotobiotic calves. The sensitivity and specificity of NCDV and Cr VP7 probes were characterized in dot blot hybridization assays, and the probes were estimated to detect at least 1 ng of viral RNA. The serotyping results obtained using VP7 probes were similar to those obtained using serologic assays. Further development of these assays may provide a useful means for the rapid detection and differentiation of bovine rotavirus serotypes in fecal samples from calves in the field.

Na-K-ATPase isoform (alpha 3, alpha 2, alpha 1) abundance in rat kidney estimated by competitive RT-PCR and ouabain binding

1996 ◽  
Vol 271 (2) ◽  
pp. F253-F260 ◽  
Author(s):  
K. Lucking ◽  
J. M. Nielsen ◽  
P. A. Pedersen ◽  
P. L. Jorgensen
Keyword(s):  
Polymerase Chain Reaction ◽  
Rat Kidney ◽  
Sodium Reabsorption ◽  
Rt Pcr ◽  
Ouabain Binding ◽  
Chain Reaction ◽  
Competitive Polymerase Chain Reaction ◽  
High Affinity ◽  
Upper Limit ◽  
Polymerase Chain

For understanding the regulation of sodium reabsorption, it is important to know whether the alpha 2- or alpha 3-isoform of Na-K-adenosinetriphosphatase (Na-K-ATPase) is expressed in mammalian kidney in addition to the predominant alpha 1 beta 1-isozyme. Here we applied competitive polymerase chain reaction (PCR) for estimation of mRNA in parenchymal zones of rat kidney for comparison to high-affinity [3H]ouabain binding. The alpha 3-isoform mRNA was demonstrated to form 0.04-0.05% of the amount of alpha 1-isoform mRNA in the cortex, medulla, and papilla of rat kidney. The alpha 2-mRNA was demonstrated in all three zones and constituted 0.03% of the amount of alpha 1-mRNA in cortex. The abundance of the alpha 1 truncated mRNA was 0.1-0.8% of that of the alpha 1-mRNA. The upper limit for expression of Na-K-ATPase isozyme with high ouabain affinity (dissociation constant, 69-141 nM) was 0.096-0.14% of the concentration of alpha 1 beta 1-Na-K-ATPase. Thus a small but well-defined pool of alpha 2- and alpha 3-isoforms constitutes < or = 0.1% of the amount of alpha 1-isoform at both the mRNA and protein level in rat kidney.

scholarly journals State of the Art and Limitations of Quantitative Polymerase Chain Reaction

2002 ◽  
Vol 85 (3) ◽  
pp. 792-796 ◽  
Author(s):  
Gordon Wiseman
Keyword(s):  
Polymerase Chain Reaction ◽  
State Of The Art ◽  
Quantitative Polymerase Chain Reaction ◽  
Processed Foods ◽  
Quantitative Real Time Pcr ◽  
Chain Reaction ◽  
Competitive Polymerase Chain Reaction ◽  
Polymerase Chain ◽  
Analytical Result ◽  
The Many

Abstract Consequential to the implementation of European Commission (EC) Regulation 1139/98, EC Regulation 49/2000, and EC Regulation 50/2000 has been the need to measure accurately the levels of the genetically modified (GM) species Roundup Ready Soya and Bt 176 Maize that are present in food. Analytical methods to detect and quantitate these transgenic species have received much attention particularly with respect to the deminimus threshold of 1% for their presence in materials derived from non-GM identity-preserved (IP) supplies. The relative advantages and limitations of threshold analysis by double-competitive polymerase chain reaction (PCR) and quantitative real-time PCR are discussed in their application to the quantitative analysis of processed foods. Consideration is also given to other factors involved in the analyses that affect the performance of quantitative procedures, and to the many uncertainties involved in the precision of a reported analytical result.

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