scholarly journals Development, Optimization, and Single Laboratory Validation of an Event-Specific Real-Time PCR Method for the Detection and Quantification of Golden Rice 2 Using a Novel Taxon-Specific Assay

2015 ◽  
Vol 63 (6) ◽  
pp. 1711-1721 ◽  
Author(s):  
Sara Jacchia ◽  
Elena Nardini ◽  
Christian Savini ◽  
Mauro Petrillo ◽  
Alexandre Angers-Loustau ◽  
...  
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Golden Rice ◽  
Specific Assay ◽  
Pcr Method ◽  
Single Laboratory ◽  
Detection And Quantification

scholarly journals Detection of Peanut Allergen Traces with a Real Time PCR Assay - The Challenge to Protect Food-Allergic Consumers

2017 ◽  
Vol 7 (1) ◽  
pp. 32 ◽  
Author(s):  
Dimitra Houhoula ◽  
Stamatios Koussissis ◽  
Vladimiros Lougovois ◽  
John Tsaknis ◽  
Dimitra Kassavita ◽  
...  
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Rapid Detection ◽  
Food Products ◽  
Molecular Techniques ◽  
Food Allergens ◽  
Pcr Assay ◽  
Peanut Allergen ◽  
Pcr Method ◽  
Detection And Quantification

The aim of the present study was the implementation of molecular techniques in the detection and quantification of allergic substances of peanut in various kinds of food products, e.g., breakfast cereals, chocolates and biscuits that are frequently related to allergies. In some cases, the presence of peanuts can be due to contamination during production and are not declared on the label. A total of 152 samples were collected from supermarkets and were analysed by a Real Time PCR method. The results indicated that 125 samples (83,3%) were found positive in peanut traces but the most important finding is that from the 84 samples that had no allergen declaration for peanuts, 48 (57,1%) of them were found positive. In conclusion, Real Time PCR can be a very important tool for the rapid detection and quantification of food allergens.

scholarly journals Application of quantitative PCR for detection of Mycoplasma suis in blood samples

2014 ◽  
Vol 58 (4) ◽  
pp. 533-539
Author(s):  
Artur Jabłoński ◽  
Dominika Borowska ◽  
Sylwia Zębek ◽  
Andrzej Kowalczyk ◽  
Arkadiusz Dors ◽  
...  
Keyword(s):  
Real Time ◽  
Quantitative Pcr ◽  
Real Time Pcr ◽  
Quantitative Method ◽  
Taqman Probe ◽  
Blood Samples ◽  
Mycoplasma Suis ◽  
Coefficients Of Variation ◽  
Pcr Method ◽  
Detection And Quantification

Abstract The aim of the study was to develop and validate a real-time PCR method, using a TaqMan probe, for quantification of Mycoplasma suis in porcine blood. No PCR signals with closely related non-haemotrophic mycoplasmas were obtained. The detection limit of PCR for plasmid combined with blood DNA was determined to be 103/reaction (5 μL of DNA) (1.2x105 target copies in 1 mL of blood). The linearity of real-time PCR (near 1) indicates its use as a quantitative method. Real-time and quantitative PCR were sensitive and specific for the detection and quantification of M. suis in the blood of animals with acute and chronic form of eperythrozoonosis. Developed quantitative PCR cannot be used to detect carrier animals with a small amount of M. suis in their blood. The validity of real-time PCR used in the studies was confirmed by the low inter- and intra-assay coefficients of variation. This fact confirms the applicability of the assay in other laboratories.

Real-time PCR method for the detection and quantification of Acanthamoeba species in various types of water samples

2013 ◽  
Vol 112 (3) ◽  
pp. 1131-1136 ◽  
Author(s):  
Po-Min Kao ◽  
Min-Che Tung ◽  
Bing-Mu Hsu ◽  
Hsien-Lung Tsai ◽  
Cheng-Yu She ◽  
...  
Keyword(s):  
Real Time ◽  
Water Samples ◽  
Real Time Pcr ◽  
Acanthamoeba Species ◽  
Pcr Method ◽  
Detection And Quantification

The importance for standardization and validation of real-time PCR for the detection of mature miR-21 in the blood of patients

2020 ◽  
pp. 66-67
Author(s):  
Л.В. Шуленина ◽  
Д.В. Салеева
Keyword(s):  
Real Time ◽  
Correlation Coefficient ◽  
Real Time Pcr ◽  
Coefficient Of Variation ◽  
Healthy Donors ◽  
Mature Micrornas ◽  
Pcr Method ◽  
Detection And Quantification

Процессы обнаружения и количественного определения методом ПЦР в реальном времени в крови пациентов зрелых микроРНК, в том числе и miR-21, должны быть стандартизированы и валидированы. Проведенные нами исследования, показывают, что специфичность метода ПЦР для miR-21 в крови здоровых доноров может быть демонстрирована образованием продукта-ампликона размером 67 нуклеотидов, прецизионность в условиях сходимости и воспроизводимости, выраженная в виде коэффициента вариации, составляет менее 2% и 3 % соответственно, а линейность метода подтверждается коэффицентом корреляции r≥0,99. The processes of detection and quantification by real-time PCR in blood of mature microRNAs, including miR-21, should be standardized and validated. Our studies show that the specificity of PCR method for miR-21 in the blood of healthy donors can be demonstrated by the formation of a 67 bp amplicon product, precision under conditions of convergence and reproducibility, expressed using the coefficient of variation, is less than 2% and 3%, respectively, and the linearity of PCR method for miR-21 is confirmed by the correlation coefficient r≥0.99.

scholarly journals TaqMan-Based Real-Time PCR Method for Detection of Yersinia pseudotuberculosis in Food

2008 ◽  
Vol 74 (20) ◽  
pp. 6465-6469 ◽  
Author(s):  
S. Thisted Lambertz ◽  
C. Nilsson ◽  
S. Hallanvuo
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Yersinia Pseudotuberculosis ◽  
Internal Amplification Control ◽  
Food Items ◽  
Specific Assay ◽  
Food Borne ◽  
Pcr Method ◽  
Probe Set

ABSTRACT A sensitive and specific assay for detection of food-borne pathogenic Yersinia pseudotuberculosis was developed. The primer-probe set was designed to target a 157-bp sequence of the chromosomally located gene ail. The complete method, including an internal amplification control, was evaluated for several different food items.

Quantitative detection and monitoring of Colletotrichum siamense in rubber trees using real-time PCR

Plant Disease ◽  
2021 ◽  
Author(s):  
Yannan Du ◽  
Meng Wang ◽  
Mingteng Long ◽  
Ye Yang ◽  
Xiaoyu Liang ◽  
...  
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Disease Risk ◽  
Rubber Tree ◽  
Detection Methods ◽  
Quantitative Detection ◽  
The Real ◽  
Colletotrichum Siamense ◽  
Pcr Method ◽  
Detection And Quantification

Colletotrichum siamense is one of the most important pathogens of rubber trees in Asia. The proper detection and quantification of C. siamense populations in rubber trees are of importance for monitoring the epidemics of the disease. In this study, we developed an ITS-based real-time PCR method to efficiently detect C. siamense infecting rubber tree, which reliably detected as little as 100 fg genomic DNA, 100 copies of target DNA and 20 conidia. The real-time PCR protocol recognized all C. siamense isolates collected from three provinces in China, while no amplification was observed with rubber tree and its other pathogens. Detection and quantification of C. siamense were performed in artificially and naturally infected rubber leaves. We could still detect C. siamense in plant mixes of which only 0.0001% of the tissue infected. An accumulation of C. siamense DNA was observed during the whole infection process at all three leaf phenological stages, suggesting the real-time PCR method can be used to monitor C. siamense development in rubber trees. Finally, the method allowed the detection of C. siamense in naturally infected and symptomless leaves of rubber trees in the fields. Compared with earlier detection methods, the real-time PCR method is more specific and more sensitive, and will be of great use for studies aiming to gain a better understanding of the epidemiology of Colletotrichum leaf disease, as well as the prediction of disease risk and the control proposal.

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