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.

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.

Detection of Sesame Seed DNA in Foods Using Real-Time PCR

2007 ◽  
Vol 70 (4) ◽  
pp. 1033-1036 ◽  
Author(s):  
JENNIFER L. BRZEZINSKI
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Sesame Seed ◽  
Food Products ◽  
Taqman Probe ◽  
2S Albumin ◽  
Sesame Seeds ◽  
Baked Goods ◽  
Pcr Method ◽  
Seed Dna

The detection of potentially allergenic foods, such as sesame seeds, in food products is a major concern for the food-processing industry. A real-time PCR method was designed to determine if sesame seed DNA is present in food products. The PCR reaction amplifies a 66-bp fragment of the sesame seed 2S albumin gene, which is detected with a sesame-specific, dual-labeled TaqMan probe. This reaction will not amplify DNA derived from other seeds present in baked goods, such as pumpkin, poppy, and sunflower seeds. Additionally, this assay will not cross-react with DNA from several tree nut species, such as almond, Brazil nut, cashew, hazelnut, and walnut, as well as four varieties of peanut. This assay is sensitive enough to detect 5 pg of purified sesame seed DNA, as well as sesame seed DNA in a spiked wheat cracker sample.

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.

Rapid Detection of Salmonella in Foods Using Real-Time PCR

2008 ◽  
Vol 71 (12) ◽  
pp. 2436-2441 ◽  
Author(s):  
CHORNG-MING CHENG ◽  
WEN LIN ◽  
KHANH THIEN VAN ◽  
LIEUCHI PHAN ◽  
NELLY N. TRAN ◽  
...  
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Rapid Detection ◽  
Food Samples ◽  
Taqman Probe ◽  
Cell Culturing ◽  
Chili Powder ◽  
Pcr Method ◽  
Food Matrices ◽  
Salmonella Serovars

Conventional methods for detection of Salmonella serovars in foods are generally time-consuming and labor intensive. A real-time PCR method has been developed with custom designed primers and a TaqMan probe to detect the presence of a 262-bp fragment of the Salmonella-specific invA gene. The method has been tested with a total of 384 field-isolated Salmonella serovars and non-Salmonella stock strains, as well as 420 U.S. Food and Drug Administration food samples, comprising a variety of food matrices. The method was highly specific in detecting Salmonella in spiked chili powder and shrimp samples, with a sensitivity of 0.04 CFU/g. In addition, the method is faster, more accurate, and less costly than the traditional U.S. Food and Drug Administration's Bacteriological Analytical Manual cell-culturing and the AOAC International–approved VIDAS methods to detect Salmonella in foods.

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