Efficient Synthesis of a Wide-Range Absorbing Azaphthalocyanine Dark Quencher and Its Application to Dual-Labeled Oligonucleotide Probes for Quantitative Real-Time Polymerase Chain Reactions

2018 ◽  
Vol 24 (38) ◽  
pp. 9658-9666 ◽  
Author(s):  
Jiri Demuth ◽  
Radim Kucera ◽  
Kamil Kopecky ◽  
Zuzana Havlínová ◽  
Antonín Libra ◽  
...  
Keyword(s):  
Real Time ◽  
Oligonucleotide Probes ◽  
Efficient Synthesis ◽  
Chain Reactions ◽  
Polymerase Chain Reactions ◽  
Wide Range ◽  
Polymerase Chain

scholarly journals Suitability of Real-Time Quantitative Polymerase Chain Reaction and Enzyme-Linked Immunosorbent Assay for cry9C Detection in Mexican Corn Tortillas: Fate of DNA and Protein After Alkaline Cooking

2004 ◽  
Vol 87 (3) ◽  
pp. 639-646 ◽  
Author(s):  
Maricarmen Quirasco ◽  
Bernd Schoel ◽  
Javier Plasencia ◽  
John Fagan ◽  
Amanda Galvez
Keyword(s):  
Real Time ◽  
Quantitative Polymerase Chain Reaction ◽  
Enzyme Linked Immunosorbent Assay ◽  
Transgenic Corn ◽  
Chain Reactions ◽  
Snack Foods ◽  
Polymerase Chain Reactions ◽  
Corn Products ◽  
Polymerase Chain ◽  
Corn Kernels

Abstract Alkaline-cooked corn, called nixtamal, is the basis for many traditional corn products such as tortillas, chips, and taco shells that are used widely in Mexico and Central America and in the preparation of snack foods that are consumed globally. To assess the effects of alkaline and thermal treatments on the detectability of DNA and protein for the presence of genetically modified sequences, various nixtamalized products were prepared from blends of conventional white corn containing 0.1, 1.0, and 10% transgenic corn (event CBH 351, StarLink™). Real-time quantitative polymerase chain reactions (RTQ–PCR) and immunoassays were used to determine the cry9C gene and protein, respectively, in unprocessed corn kernels, freshly prepared alkaline-cooked and ground corn (masa), masa flour, tortillas prepared from masa by heat treatment, chips prepared from damp masa dough by deep frying, and from tortillas processed at high (200°C) and low temperatures (70°C). In spite of progressive degradation of genomic DNA during processing, RTQ–PCR genetic analysis allowed detection and quantification of the cry9C gene in all products prepared from 10, 1, and 0.1% StarLink corn, except deep-fried chips containing 0.1% StarLink. Enzyme-linked immunosorbent assays readily detected <1ppm cry9C protein in all blends of unprocessed corn (10, 1, and 0.1% StarLink) as well as in nonfried tortilla and masa products. This technique was not suitable for thermally treated nixtamalized products containing <1% transgenic corn.

Parallel detection of five human herpes virus DNAs by a set of real-time polymerase chain reactions in a single run

2003 ◽  
Vol 26 (1) ◽  
pp. 85-93 ◽  
Author(s):  
Markus Stöcher ◽  
Victoria Leb ◽  
Michael Bozic ◽  
Harald H Kessler ◽  
Gabriele Halwachs-Baumann ◽  
...  
Keyword(s):  
Real Time ◽  
Herpes Virus ◽  
Human Herpes ◽  
Chain Reactions ◽  
Parallel Detection ◽  
Polymerase Chain Reactions ◽  
Human Herpes Virus ◽  
Polymerase Chain ◽  
Herpès Virus

scholarly journals Detection of Bovine Leukemia Virus in Blood and Milk by Nested and Real-Time Polymerase Chain Reactions

2003 ◽  
Vol 15 (1) ◽  
pp. 72-76 ◽  
Author(s):  
Christopher J. Kuckleburg ◽  
Christopher C. Chase ◽  
Eric A. Nelson ◽  
Salvatore A. E. Marras ◽  
Matthew A. Dammen ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Early Detection ◽  
Real Time ◽  
Bovine Leukemia Virus ◽  
Leukemia Virus ◽  
Chain Reaction ◽  
Nested Polymerase Chain Reaction ◽  
Chain Reactions ◽  
Polymerase Chain Reactions ◽  
Polymerase Chain

Concerns about retroviruses in livestock and products derived from them have necessitated the development of tests to detect the bovine leukemia virus (BLV) in blood and milk from cattle. Dairy cattle ( n = 101) from 5 different geographical areas were used for this study. A nested polymerase chain reaction (PCR) identified 98% of BLV seropositive cattle ( n = 80) from blood and 65% from milk, whereas real-time PCR detected 94% of BLV seropositive cattle from blood and 59% from milk. Bovine leukemia virus was also detected by PCR in approximately 10% of seronegative cattle ( n = 21), most likely because of early detection before seroconversion.

scholarly journals Decreased Expression of Hsa_circ_00001649 in Gastric Cancer and Its Clinical Significance

2017 ◽  
Vol 2017 ◽  
pp. 1-6 ◽  
Author(s):  
Wen-han Li ◽  
Yong-chun Song ◽  
Hao Zhang ◽  
Zhang-jian Zhou ◽  
Xin Xie ◽  
...  
Keyword(s):  
Gastric Cancer ◽  
Expression Level ◽  
Serum Samples ◽  
Chain Reactions ◽  
Normal Tissues ◽  
Polymerase Chain Reactions ◽  
Potential Biomarker ◽  
Wide Range ◽  
Pathological Differentiation ◽  
Polymerase Chain

Background. It has been reported that circRNAs are differentially expressed in a wide range of cancers and could be used as a new biomarker for diagnosis. However, the correlation between circRNAs and gastric cancer (GC) it is still unclear. Materials and Methods. In this study, by using real-time quantitative reverse transcription-polymerase chain reactions (qRT-PCRs), we detected the expression level of hsa_circ_0001649 in tissue and serum samples from GC patients. Results. We found that hsa_circ_0001649 expression was significantly downregulated in GC tissue compared with their paired paracancerous histological normal tissues (PCHNTs) (P<0.01). We next analyzed the expression level of hsa_circ_0001649 in serum samples between preoperative and postoperative GC patients. We found that its level in serum was significantly upregulated after surgery (P<0.01). The area under the receiver operating characteristic (ROC) curve was 0.834. Moreover, the expression level of hsa_circ_0001649 was significantly correlated with pathological differentiation (P=0.039). Conclusion. Our test suggested that hsa_circ_0001649 was significantly downregulated in GC and may become a novel potential biomarker in the diagnosis of GC.

An empirical approach to target DNA quantification in environmental samples using real-time polymerase chain reactions

2007 ◽  
Vol 39 (8) ◽  
pp. 1956-1967 ◽  
Author(s):  
Robert H. Gulden ◽  
David Levy-Booth ◽  
Rachel Campbell ◽  
Jeff R. Powell ◽  
Miranda M. Hart ◽  
...  
Keyword(s):  
Real Time ◽  
Environmental Samples ◽  
Dna Quantification ◽  
Empirical Approach ◽  
Chain Reactions ◽  
Polymerase Chain Reactions ◽  
Target Dna ◽  
Polymerase Chain

Evaluation of the pooling of swabs for real-time PCR detection of low titre shedding of low pathogenicity avian influenza in turkeys

2012 ◽  
Vol 141 (6) ◽  
pp. 1286-1297 ◽  
Author(s):  
M. E. ARNOLD ◽  
M. J. SLOMKA ◽  
V. J. COWARD ◽  
S. MAHMOOD ◽  
P. J. RALEIGH ◽  
...  
Keyword(s):  
Avian Influenza ◽  
Real Time ◽  
Detrimental Effect ◽  
Bayesian Modelling ◽  
Chain Reactions ◽  
Polymerase Chain Reactions ◽  
Buccal Swabs ◽  
Detection Window ◽  
Polymerase Chain ◽  
Turkey Flock

SUMMARYThe purpose of this study was to determine whether pooling avian influenza (AI)-positive swabs with negative swabs has a detrimental effect on the sensitivity of AI real-time reverse transcription–polymerase chain reactions (rRT–PCRs). Cloacal and buccal swabs were sampled daily from 12 turkeys infected with A/goose/England/07(H2N2). For half the turkeys, each swab was mixed with four swabs from known AI-negative turkeys, and for the other half the swabs were tested individually. Bayesian modelling was used to (i) determine whether pooling the positive swabs compromised the cycle threshold (Ct) value obtained from the rRT–PCRs, and (ii) estimate the likelihood of detection of an H2N2 infected turkey flock via rRT–PCR for pooled and individually tested swabs (cloacal and buccal)vs. the number of days post-infection of the flock. Results indicated that there was no significant effect of compromising AI rRT–PCR sensitivity by pooling a weak positive swab with negative swabs on the Ctvalues which were obtained. Pooled sampling was able to widen the detection window compared to individual sampling, for the same number of rRT–PCR tests. This indicates that pooled sampling would be an effective method of reducing the number of tests to be performed to determine flock status during an AI outbreak and for surveillance.

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