Using Real-Time PCR as a tool for monitoring the authenticity of commercial coffees

Ferreira, Thiago; Farah, Adriana; Oliveira, Tereza Cristina Souza de; Lima, Ivanilda S.; Vitório, Felipe; Oliveira, Edna Maria Moraes

doi:10.1016/j.foodchem.2015.12.045

Cited by 45 publications

(26 citation statements)

References 19 publications

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“…The LOD was set to the minimum value detected above 95% of analysis results (<5% false negative result). The LOQ was determined to be a value of MCq − (2 × SD) below 35 [17,18].…”

Section: Qualitative and Quantitative Analysis By Multiplex Real-timementioning

confidence: 99%

Development of multiplex real-time PCR for rapid identification and quantitative analysis of Aspergillus species

Kim¹,

Park²,

Cho³

et al. 2020

PLoS ONE

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The identification of Aspergillus species and azole resistance is highly important for the treatment of invasive aspergillosis (IA), which requires improvements in current fungal diagnostic methods. We aimed to develop multiplex real-time PCR to identify major Aspergillus section and azole resistance. BenA and cyp51A genes were used to design primers, probes, and control DNA for multiplex PCR. Qualitative and quantitative analysis was conducted for 71 Aspergillus and 47 non-Aspergillus isolates. Further, the limit of detection (LOD) and limit of quantitation (LOQ) from hyphae or conidia were determined according to the culture time. Newly developed real-time PCR showed 100% specificity to each Aspergillus section (Fumigati, Nigri, Flavi, and Terrei), without cross-reaction between different sections. In quantitative analysis of sensitivity measurements, LOD and LOQ were 40 fg and 400 fg, respectively. Melting temperature analysis of the cyp51A promoter to identify azole resistance showed temperatures of 83.0 ± 0.3˚C and 85.6 ± 0.6˚C for susceptible A. fumigatus and resistant isolates with TR34 mutation, respectively. The minimum culture time and fungal colony size required for successful detection were 24 h and 0.4 cm in diameter, respectively. The developed multiplex real-time PCR can identify common Aspergillus sections quantitatively and detect presence of the TR34 mutation. Further, this method shows high sensitivity and specificity, allowing successful detection of early-stage fungal colonies within a day of incubation. These results can provide a template for rapid and accurate diagnosis of IA.

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“…The LOD was set to the minimum value detected above 95% of analysis results (<5% false negative result). The LOQ was determined to be a value of MCq − (2 × SD) below 35 [17,18].…”

Section: Qualitative and Quantitative Analysis By Multiplex Real-timementioning

confidence: 99%

Development of multiplex real-time PCR for rapid identification and quantitative analysis of Aspergillus species

Kim¹,

Park²,

Cho³

et al. 2020

PLoS ONE

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“…Brazil is the largest coffee producer in the world. In this sector, the main fraud reported was the addition of barley, corn, rice, and coffee husk (Ferreira et al., ; Moura Ribeiro, Boralle, Pezza, Pezza, & Toci, ). The fraudulent addition of sucrose (Mendes, Quadri & Quadri, ), soil, and sand (Santos & Abrantes, ) to tea was also reported.…”

Section: Discussionmentioning

confidence: 99%

Economically Motivated Food Fraud and Adulteration in Brazil: Incidents and Alternatives to Minimize Occurrence

Tibola

Silva

Dossa

et al. 2018

Journal of Food Science

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“…The amplification efficiency was calculated on the basis of the standard curve using the equations E = 10 −1 /slope, and efficiency (%) = (E −1 ) × 100. The LOD was regarded as the analytical concentration at which the method detected the presence of a target gene in at least 95% of true-positive biological samples (< 5% of false-negative results) [22].…”

Section: Determination Of Amplification Efficiency Correlation Coeffmentioning

confidence: 99%

Development of DNA-based species-specific real-time PCR markers for four berry fruits and their application in commercial berry fruit foods

Moon

Kim

et al. 2019

Appl Biol Chem

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Berry fruits have attracted attention because of their purported benefits for aging, cardiovascular disease, and cancer. Therefore, highly priced berry fruits might be targets for food adulteration and fraud. In this study, eight speciesspecific primer sets based on the single nucleotide polymorphism of the chloroplast genomes of four berry fruits (aronia, blackberry, cranberry, and strawberry) were developed for quantitative real-time PCR (qPCR) analysis by SYBR Green staining with the aim of preventing berry fruit food fraud. The developed primer pairs exhibited high efficiencies ranging from 88 to 110% with strong correlation coefficients (R 2 > 0.99) for the amplification of each target species. However, no clear correlation coefficients were evident for non-target species. To evaluate the practicality of the developed primers, 18 commercial berry fruit products were verified by qPCR analysis. The developed primer pairs were amplified to a low C t value (range 16.1-23.3) for the target species and proved capable of detecting target species in berry fruit commercial foods. Therefore, the developed qPCR-based species-specific markers could be suitable for the prevention of berry fruit food fraud and to verify marker reliability. which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made.

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Using Real-Time PCR as a tool for monitoring the authenticity of commercial coffees

Cited by 45 publications

References 19 publications

Development of multiplex real-time PCR for rapid identification and quantitative analysis of Aspergillus species

Development of multiplex real-time PCR for rapid identification and quantitative analysis of Aspergillus species

Economically Motivated Food Fraud and Adulteration in Brazil: Incidents and Alternatives to Minimize Occurrence

Development of DNA-based species-specific real-time PCR markers for four berry fruits and their application in commercial berry fruit foods

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