Evaluation of locked nucleic acid and TaqMan probes for specific detection of cashew nut in processed food by real time PCR

Sanchiz, África; Ballesteros, Isabel; Marques, Eric de Lima Silva; Diéguez, M.; Rueda, Julia; Cuadrado, Carmen; Linacero, Rosario

doi:10.1016/j.foodcont.2018.02.021

Cited by 16 publications

(26 citation statements)

References 33 publications

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“…The quantity and quality of the extracted DNA were evaluated on 0.8% TBE agarose gels and using a Nanodrop ND-1000 spectrophotometer (Thermo-Fisher, Waltham, MA, USA), taking into account the values obtained by measuring absorbance at 230, 260 and 280 nm. We also used DNA obtained in our previous studies [20][21][22], from other sequenced varieties of hazelnut (Pauetet, Tonda and Sant Giovanni), pistachio (Aegina, Mateur and Sirora) and cashew (Embrapa 50, BRS 189, BRS 274 and CCP06).…”

Section: Dna Extraction and Conventional Pcrmentioning

confidence: 99%

“…Positive amplification of all samples was tested by PCR using universal eukaryotic primers for 18S described elsewhere (forward 5 -CGCGAGAAGTCCACTAAACC-3 , reverse 5 -CCTACGGAAACCTTGTTACGA-3 ) [22].…”

Section: Dna Extraction and Conventional Pcrmentioning

confidence: 99%

“…Primers were designed in allergen-coding regions of hazelnut, pistachio and cashew, being conserved among the different varieties ( Figure S1), and targets were selected in the Cor a 9, Pis v 1 and Ana o 1 allergen-coding sequences, respectively [20][21][22]. After that, the performance of primers and probes was assayed, regarding specificity, sensitivity and reaction efficiency.…”

Section: Targets Primer Design and Real-time Pcr Conditionsmentioning

confidence: 99%

“…For the study we present here, a hydrolysis probe has been designed in order to standardize the rest of the assays and as an attempt to improve the sensitivity of the method. In pistachio and cashew, Real-Time PCR-based detection methods were recently published by our group [21,22], selecting Pis v 1 and Ana o 1 as target. In these cases, a Locked Nucleic Acid (LNA, Roche, Basel, Switzerland) probe and a hydrolysis probe were designed and used for pistachio and cashew detection, respectively.…”

Section: Targets Primer Design and Real-time Pcr Conditionsmentioning

confidence: 99%

“…Before this study, the development and validation of Real-Time PCR methods for pistachio, cashew and hazelnut were performed and published [20][21][22]. In the three works, allergen-coding genes were selected as targets (Pis v 1 for pistachio, Ana o 1 for cashew and Cor a 9 for hazelnut).…”

Section: Introductionmentioning

confidence: 99%

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Effect of Instant Controlled Pressure Drop (DIC) Treatment on the Detection of Nut Allergens by Real Time PCR

Sanchiz

Cuadrado

Haddad

et al. 2020

Foods

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Tree nuts show nutritional properties and human health benefits. However, they contain allergenic proteins, which make them harmful to the sensitised population. The presence of tree nuts on food labelling is mandatory and, consequently, the development of suitable analytical methodologies to detect nuts in processed foods is advisable. Real-Time PCR allowed a specific and accurate amplification of allergen sequences. Some food processing methods could induce structural and/or conformational changes in proteins by altering their allergenic capacity, as well as produce the fragmentation and/or degradation of genomic DNA. In this work, we analysed by means of Real-Time PCR, the influence of pressure and thermal processing through Instant Controlled Pressure Drop (DIC) on the detectability of hazelnut, pistachio and cashew allergens. The detection of targets in hazelnut, pistachio and cashew (Cor a 9, Pis v 1 and Ana o 1, respectively) is affected by the treatment to different extents depending on the tree nut. Results are compared to those previously obtained by our group in the analysis of different treatments on the amplificability of the same targets. Reduction in amplificability is similar to that reported for some autoclave conditions. Our assays might allow for the detection of up to 1000 mg/kg of hazelnut, pistachio and cashew flours after being submitted to DIC treatment in food matrices.

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Section: Dna Extraction and Conventional Pcrmentioning

confidence: 99%

Section: Dna Extraction and Conventional Pcrmentioning

confidence: 99%