Multiplex qPCR assay for simultaneous quantification of CYP51‐S524T and SdhC‐H152R substitutions in European populations of Zymoseptoria tritici

2020 ◽  
Vol 69 (9) ◽  
pp. 1666-1677
Author(s):  
Pierre Hellin ◽  
Maxime Duvivier ◽  
Aurélie Clinckemaillie ◽  
Charlotte Bataille ◽  
Anne Legrève ◽  
...  
Keyword(s):  
Qpcr Assay ◽  
Zymoseptoria Tritici ◽  
Multiplex Qpcr ◽  
Simultaneous Quantification ◽  
European Populations

scholarly journals Evaluation of a fully automated high-throughput SARS-CoV-2 multiplex qPCR assay with built-in screening functionality for del-HV69/70- and N501Y variants such as B.1.1.7

2021 ◽  
pp. 104894
Author(s):  
Dominik Nörz ◽  
Moritz Grunwald ◽  
Flaminia Olearo ◽  
Nicole Fischer ◽  
Martin Aepfelbacher ◽  
...  
Keyword(s):  
High Throughput ◽  
Qpcr Assay ◽  
Multiplex Qpcr

scholarly journals Multiplex Real-Time Polymerase Chain Reaction for Simultaneous Quantification ofSalmonellaspp.,Escherichia coli, andStaphylococcus aureusin Different Food Matrices: Advantages and Disadvantages

2018 ◽  
Vol 2018 ◽  
pp. 1-12 ◽  
Author(s):  
Amanda Teixeira Sampaio Lopes ◽  
George Rêgo Albuquerque ◽  
Bianca Mendes Maciel
Keyword(s):  
Escherichia Coli ◽  
Real Time ◽  
Limit Of Detection ◽  
Microbiological Quality ◽  
Rapid Screening ◽  
Multiplex Qpcr ◽  
Advantages And Disadvantages ◽  
Simultaneous Quantification ◽  
Polymerase Chain ◽  
Food Matrices

Quantitative real-time polymerase chain reactions (qPCRs) of the most prevalent bacteria causing foodborne diseases worldwide, such asSalmonellaspp.,Escherichia coli, andStaphylococcus aureus,can be an important tool for quantitative microbial risk assessment, which requires numerical data to determine the level of contamination at a specific stage of food production. However, most of qPCR assays described in the literature for these pathogens are qualitative; their objective is pathogen detection and not pathogen quantification. Thus, the aim of our work was to develop a qPCR for the simultaneous quantification ofSalmonellaspp.,E. coli, andS. aureusand to propose its use in the analysis of foods, as a tool for microbiological quality monitoring. For this, a multiplex qPCR was standardized for the simultaneous quantification of specific fragments of target genes (ssf,phoA, andnuc) corresponding to each one of the mentioned bacteria. The limit of detection of the technique was 13, 10, and 12 gene copies forssf,phoA, andnuc, respectively; standard curves showed R2> 0.99, with efficiencies ranging from 99 to 110%, and inter- and intraexperiment reproducibility presented a low coefficient of variation in all trials. This methodology was applied in different food matrices (milk, ground beef, and oyster meat), and the results were compared with official microbiological culture methodology and with ready-to-use test. Advantages and disadvantages of each methodology used in this study are pointed out. We suggest that this multiplex qPCR can be used as a rapid screening technique for the analysis of food microbiological quality.

Multiplex qPCR assay for assessment of reference gene expression stability in rat tissues/samples

2020 ◽  
Vol 53 ◽  
pp. 101611 ◽  
Author(s):  
Alexander P. Schwarz ◽  
Daria A. Malygina ◽  
Anna A. Kovalenko ◽  
Alexander N. Trofimov ◽  
Aleksey V. Zaitsev
Keyword(s):  
Gene Expression ◽  
Reference Gene ◽  
Qpcr Assay ◽  
Rat Tissues ◽  
Expression Stability ◽  
Gene Expression Stability ◽  
Multiplex Qpcr ◽  
Reference Gene Expression

scholarly journals Field application of a novel multiplex qPCR assay reveals the occurrence of the zoonotic hookworm Ancylostoma braziliense in Nigerian dogs

Acta Tropica ◽  
2021 ◽  
Vol 213 ◽  
pp. 105758
Author(s):  
Luca Massetti ◽  
Joshua Kamani ◽  
Anke Wiethoelter ◽  
Phillip McDonagh ◽  
Vito Colella ◽  
...  
Keyword(s):  
Field Application ◽  
Qpcr Assay ◽  
Multiplex Qpcr

scholarly journals Multiplex qPCR Assay for Direct Detection and Quantification of Colletotrichum truncatum, Corynespora cassiicola, and Sclerotinia sclerotiorum in Soybean Seeds

Plant Disease ◽  
2020 ◽  
Vol 104 (11) ◽  
pp. 3002-3009
Author(s):  
Maísa Ciampi-Guillardi ◽  
Juliana Ramiro ◽  
Maria Heloisa Duarte de Moraes ◽  
Marina Coan Goldoni Barbieri ◽  
Nelson S. Massola
Keyword(s):  
Sclerotinia Sclerotiorum ◽  
Internal Control ◽  
Direct Detection ◽  
Simultaneous Detection ◽  
Plant Diseases ◽  
Qpcr Assay ◽  
Soybean Seeds ◽  
Corynespora Cassiicola ◽  
Colletotrichum Truncatum ◽  
Multiplex Qpcr

Precise diagnosis of plant diseases is one of the most effective tools to minimize yield losses. Colletotrichum truncatum, Corynespora cassiicola, and Sclerotinia sclerotiorum are common soilborne pathogens that affect soybeans all over the world. We developed a multiplex quantitative real-time polymerase chain reaction (qPCR) assay to simultaneously detect and quantify the three pathogens in soybean seeds and to survey their occurrence in the main soybean production areas in Brazil. Species-specific primers and probes for C. truncatum and C. cassiicola were designed based on GAPDH and TEF1 genes, respectively, to be combined with qPCR detection of S. sclerotiorum previously reported. The multiplex qPCR assay was successful in the simultaneous detection of C. truncatum, C. cassiicola, and S. sclerotiorum, along with a host internal control. The four pathogens were detected and quantified in artificially and naturally infested soybean seeds, even in the lowest incidence level tested of 0.0625% or 1 infected seed out of 1,599 healthy ones. From 81 seed samples tested, C. truncatum was the most frequently detected pathogen and with higher incidence levels (0.25 to 0.125%), followed by S. sclerotiorum and C. cassiicola, both with lower incidence levels (0.125 to 0.0625%). Together, the results evidenced the high sensitivity of the multiplex qPCR assay, indicating its usefulness for a quick and reliable diagnosis of soybean diseases in seeds.

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