Development of ultrasensitive direct chemiluminescent enzyme immunoassay for determination of aflatoxin M1 in milk

Vdovenko, Marina M.; Lu, Chuan-Chen; Yu, Feng‐Yih; Sakharov, Ivan Yu.

doi:10.1016/j.foodchem.2014.02.128

Cited by 65 publications

(28 citation statements)

References 23 publications

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“…Based on this, in the past years, a series of modified ELISA methods have appeared. For example, Vdovenko et al (2014) developed a competitive immunological assay of chemiluminescence (CL ELISA) for detection of AFM 1 . To improve the method's sensitivity, a mixture of 3-(10-phenotiazine)-propane-1sulphonate (SPTZ) and 4-morpholinopyridine (MORPH) was used for increasing the induction of peroxidase.…”

Section: Enzyme-linked Immunosorbent Assaymentioning

confidence: 99%

Recent aflatoxin survey data in milk and milk products: A review

Ketney

Santini

Oancea

2017

Int J of Dairy Tech

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Aflatoxin M1 (AFM1) occurrence in human and animal milk, infant formula, powdered milk, cheese and yoghurt represents a risk for health. The last four years (2010–2014) of data, as well as the most frequently and updated analytical methods applied for AFM1 quantification, are evaluated. Aflatoxin B1, considered the most potent toxic aflatoxin, is metabolised to form the monohydroxy derivative AFM1. This metabolized, expressed in the milk, is relatively stable, and it is not eliminated by heat treatments or pasteurisation, and thus represents a serious health concern.

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Section: Enzyme-linked Immunosorbent Assaymentioning

confidence: 99%

Recent aflatoxin survey data in milk and milk products: A review

Ketney

Santini

Oancea

2017

Int J of Dairy Tech

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“…8,14,15 However, the quantitative approaches require complicated pretreatment, professional operators and expensive instruments. Some immunological methods like enzyme-linked immune sorbent assay (ELISA) [16][17][18] and immunosensors [19][20][21] have also been reported for AFM 1 detection. There are disadvantages in both preparation of the antibody and its stability and this limits its application in the field.…”

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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“…Regarding the method used for the identification of AFM 1 in bovine milk, it showed a good sensitivity and precision for the quantification of aflatoxin within the Brazilian parameters, although there are also very sensitive methods for detection of aflatoxin M1 (Vdovenko et al, 2014;Kanungo et al, 2011).…”

Section: Resultsmentioning

confidence: 99%

Determination of aflatoxin M1 in bovine milk from the Alagoas/Brazil State dairy belt by high performance liquid chromatography (HPLC)

Alves¹,

Lima²,

Costa³

et al. 2018

Afr. J. Microbiol. Res.

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Milk is considered a nutritionally noble food and is therefore suitable for feeding children and adults. However, contamination of milk by mycotoxins may pose a health risk to the consumer. Aflatoxins, mycotoxins produced by fungi of the genus Aspergillus, can be found in several food products, including milk and its derivatives, which reinforces the importance of this type of study on the occurrence of the aflatoxin M 1 (AFM 1 ) in raw milk. We analyzed 45 samples of raw bovine milk from expansion tanks from 23 farms in different municipalities of the dairy belt of the state of Alagoas/Brazil. Samples were collected directly from the cooling tanks and transported under refrigeration for analysis. The method used for the extraction of AFM 1 was that proposed by the Adolfo Lutz Institute. On the other hand, the detection of AFM 1 occurred by high performance liquid chromatography (HPLC) by identifying retention times. The results of the analyses indicated that none of the collected samples presented contamination by aflatoxin M 1 , thus indicating that the milk commercialized in Alagoas shows a good quality against this toxic agent.

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Development of ultrasensitive direct chemiluminescent enzyme immunoassay for determination of aflatoxin M1 in milk

Cited by 65 publications

References 23 publications

Recent aflatoxin survey data in milk and milk products: A review

Recent aflatoxin survey data in milk and milk products: A review

A qPCR aptasensor for sensitive detection of aflatoxin M1

Determination of aflatoxin M1 in bovine milk from the Alagoas/Brazil State dairy belt by high performance liquid chromatography (HPLC)

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