Development of an impedimetric aptasensor for the determination of aflatoxin M1 in milk

Istamboulié, Georges; Paniel, Nathalie; Zara, Lorena; Granados, Lucia Reguillo; Barthelmebs, Lise; Noguer, Thierry

doi:10.1016/j.talanta.2015.09.012

Cited by 124 publications

(54 citation statements)

References 27 publications

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“…Aptasensors were developed for the detection of AFM 1 using electrochemical methods and impedance spectroscopy techniques. 30,31 However, the 4 selectivity of the aptamer for AFM 1 was not clear since only irrelevant OTA was chosen as an interference to study the cross-reaction. Cross-activity tests between other toxins (AFB 1 , AFB 2 , AFG 1 and AFG 2 ) should be carried out to answer the question whether the aptasensor is suitable for quantifying the AFM 1 concentration in real samples.…”

Section: Introductionmentioning

confidence: 99%

A qPCR aptasensor for sensitive detection of aflatoxin M1

et al. 2016

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. In addition, the aptasensor developed here exhibits high selectivity for AFM 1 over other mycotoxins and small effects from cross-reaction with structural analogs. The method proposed here has been successfully applied to quantitative determination of AFM 1 in infant rice cereal and infant milk powder samples. Results demonstrated that the current approach is potentially useful for food safety analysis, and it could be extended to a large number of targets. IntroductionAflatoxin M 1 (AFM 1 ), one of the most toxic contaminants in dairy products, is a metabolite produced by dairy cows as a result of being fed with feeds contaminated with Aflatoxin B 1 (AFB 1 ). [3][4][5] Once present in dairy products, AFM 1 poses a hazard to humans (especially infants) who consume them. Results and Discussion Optimization of the amplification of complementary ssDNAThe complementary ssDNA is applied as the subsequent PCR template. The Optimization of streptavidin and the biotinylated aptamerOther factors would affect the performance of this method including the concentrations of streptavidin, the biotinylated aptamer and the complementary ssDNA and these all should be optimized. Under a fixed concentration of complementary ssDNA at 10 nM, the concentrations of streptavidin and the 6 biotinylated aptamer were analyzed for the PCR amplification signal change (Fig. S3).The adsorptive power of streptavidin to PCR tubes and the binding ability of streptavidin-coated tubes to biotinylated aptamer was primarily detected using different concentrations of streptavidin (Fig. S3). This also shows that there was a clear difference of Ct values between the control group and streptavidin-coated tubes, which showed the intense adsorptive ability between biotinylated aptamer and streptavidin-coated tubes. In analysis of the Ct values at different concentrations of streptavidin, the Ct value reached the lowest level when the concentration of streptavidin was 2.5 ng mL -1 . This graph also shows that the Ct values decreased with the increase of the amount of aptamer when aptamer levels were below 10.0 nM andalso that the Ct values increased with the amount of aptamer above 10.0 nM mainly because of steric hindrance. Thus, 2.5 ng mL -1 of streptavidin and 10 nM of aptamer were the optimal conditions for RT-qPCR amplification. AFM 1 determinationUnder optimal conditions, the amplification curves and calibration curve of this aptasensor for different levels of AFM 1 with AFM 1 DNA1 were determined by RT-qPCR (Fig. 1). As is shown in Fig. 1(A), the cycle number increased with an increase in AFM 1 . More complementary ssDNA would be released when more AFM 1 is present in the reaction system, which leads to a decrease in the amount of the PCR template and an increase in cycle number. Meanwhile, a good linear relationshipbetween Ct values and AFM 1 levels in the range of 1×10 -4 to 1 µg L -1 was obtained as (Table 1). Specificity analysisThe specificity of the aptasensor plays an important role in the development and practicality of this method. In ...

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Section: Introductionmentioning

confidence: 99%

A qPCR aptasensor for sensitive detection of aflatoxin M1

et al. 2016

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“…Using impedance spectroscopy, a detection limit of 1-14 ng/mL was obtained. Very recently, an impedimetric aptasensor for detection of AFM1 was reported by Istamboulie et al [78]. In this work, the hexaethyleneglycol-modified oligonucleotides (seven base pair sequences) of anti-AFM1 aptamer were immobilized on the diazotized screen-printed carbon electrode (SPCEs) via a carbodiimide coupling reaction.…”

Section: Electrochemical Aptasensors For Afs Detectionmentioning

confidence: 99%

Recent Advances in Electrochemical-Based Sensing Platforms for Aflatoxins Detection

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Abstract:Mycotoxin are small (MW~700 Da), toxic secondary metabolites produced by fungal species that readily colonize crops and contaminate them at both pre-and post-harvesting. Among all, aflatoxins (AFs) are mycotoxins of major significance due to their presence in common food commodities and the potential threat to human health worldwide. Based on the severity of illness and increased incidences of AFs poisoning, a broad range of conventional and analytical detection techniques that could be useful and practical have already been reported. However, due to the variety of structural analogous of these toxins, it is impossible to use one common technique for their analysis. Numerous recent research efforts have been directed to explore alternative detection technologies. Recently, immunosensors and aptasensors have gained promising potential in the area of sample preparation and detection systems. These sensors offer the advantages of disposability, portability, miniaturization, and on-site analysis. In a typical design of an aptasensor, an aptamer (ssDNA or RNA) is used as a bio-recognition element either integrated within or in intimate association with the transducer surface. This review paper is focused on the recent advances in electrochemical immunoand aptasensing platforms for detection of AFs in real samples.

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“…In this work, the aptamer, modified with hexaethyleneglycol, was covalently immobilized on a SPCE surface activated with diazonium salts. The method was successfully applied toreal milk samples with a good correlation with a conventional immunoassay [83]. Using the same immobilization strategy, we described an impedimetric aptasensor for AFB1 by comparing two different aptamers.…”

Section: Label-free Aptasensors For Myctotoxin Determinationmentioning

confidence: 99%

Label-Free Aptasensors for the Detection of Mycotoxins

Rhouati

Catanante

Nunes

et al. 2016

Sensors

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Various methodologies have been reported in the literature for the qualitative and quantitative monitoring of mycotoxins in food and feed samples. Based on their enhanced specificity, selectivity and versatility, bio-affinity assays have inspired many researchers to develop sensors by exploring bio-recognition phenomena. However, a significant problem in the fabrication of these devices is that most of the biomolecules do not generate an easily measurable signal upon binding to the target analytes, and signal-generating labels are required to perform the measurements. In this context, aptamers have been emerged as a potential and attractive bio-recognition element to design label-free aptasensors for various target analytes. Contrary to other bioreceptor-based approaches, the aptamer-based assays rely on antigen binding-induced conformational changes or oligomerization states rather than binding-assisted changes in adsorbed mass or charge. This review will focus on current designs in label-free conformational switchable design strategies, with a particular focus on applications in the detection of mycotoxins.

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Development of an impedimetric aptasensor for the determination of aflatoxin M1 in milk

Cited by 124 publications

References 27 publications

A qPCR aptasensor for sensitive detection of aflatoxin M1

A qPCR aptasensor for sensitive detection of aflatoxin M1

Recent Advances in Electrochemical-Based Sensing Platforms for Aflatoxins Detection

Label-Free Aptasensors for the Detection of Mycotoxins

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