Development of nanocolloidal gold based immunochromatographic assay for rapid detection of transgenic vegetative insecticidal protein in genetically modified crops

Kumar, Rajesh; Singh, Chandra K.; Kamle, Suchitra; Sinha, Rajeshwar P.; Bhatnagar, Raj K.; Kachru, D.N.

doi:10.1016/j.foodchem.2010.03.086

Cited by 36 publications

(29 citation statements)

References 17 publications

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“…The level of detection of proteins obtained in this study at contamination levels of up to 1% corroborates et al, 2012;CUNHA et al, 2005;BULCKE et al, 2007;KUMAR et al, 2010;KUMAR, 2012). This shows the efficiency and repeatability of the results, even when using different transgenic events and crops.…”

Section: Immunochromatographic Strip Test (Lateral Flow Membrane Strisupporting

confidence: 89%

Detection of transgenic events in maize using immunochromatographic strip test and conventional PCR

Cantelmo

Pinho

et al. 2013

Ciênc. agrotec.

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With the growth in the transgenic market, fast and economically viable methodologies are necessary for undertaking transgene detection tests, both for identification of contamination in seeds and in grain. Seeds from commercial conventional GNZ 2004, and transgenic VT-Pro (MON89034), Roundup Ready (NK603) and Herculex (TC1507) maize cultivars were used. In order to simulate different levels of contamination, the transgenic seeds were mixed with conventional seeds at levels of 0.2%, 0.4%, 1.0% and 1.6% for VT-Pro, and 0.2%, 0.5%, 0.8% and 1.2% for Roundup Ready and Herculex. The lateral flow membrane strip test was performed in the whole seed, endosperm and embryo. For evaluation of the specificity of the technique in detection of the TC1507 event by means of the conventional PCR technique, seeds of the commercial maize hybrid GNZ 2004 were used as negative control, and the maize hybrid 2B655Hx as positive control. In order to simulate different levels of contamination, transgenic seeds were mixed with conventional seeds at the levels of 10%, 5%, 1%, 0.5% and 0.1%. Seeds from each sample were crushed, and then DNA extraction was performed by the CTAB 2% method. Using the immunochromatographic strip, it was possible to evaluate the expression of proteins related to the VT-Pro, Roundup Ready and Herculex events when whole seeds were used at the 0.2% level of contamination, whereas by the conventional PCR technique, it was possible to detect the TC1507 event in samples with 1% contamination.Index terms: Fast testing, contamination, specificity, transgenesis. RESUMOCom o crescimento do mercado de transgênicos, fazem-se necessárias metodologias rápidas e economicamente viáveis para detecção de transgenes, tanto para a identificação de contaminação em sementes quanto em grãos. Foram utilizadas sementes de cultivares comerciais de milho GNZ 2004 convencional, VT-Pro (MON89034), Round up Ready -RR (NK603) e Herculex (TC1507). Afim de simular diferentes níveis de contaminação, sementes transgênicas foram misturadas às sementes convencionais, nos níveis de 0,2%, 0,4%, 1,0% e 1,6%, para VT-Pro e 0,2%, 0,5%, 0,8% e 1,2%, para RR e Herculex. O teste de tiras foi realizado em semente inteira, endosperma e embrião. Para a avaliação da especificidade da técnica na detecção do evento TC1507, por meio da técnica de PCR convencional, foram utilizadas, como controle negativo, sementes do híbrido comercial de milho GNZ 2004 e, como controle positivo, o híbrido de milho 2B655Hx. Com a finalidade de simular diferentes níveis de contaminação, sementes transgênicas (TC1507) foram misturadas às sementes convencionais, nos níveis de 10%, 5%, 1%, 0,5% e 0,1%. Sementes de cada amostra foram trituradas e foi realizada a extração do DNA pelo método CTAB 2%. Utilizando-se a tira imunocromatográfica, foi possível avaliar a expressão de proteínas referente aos eventos VT-Pro, RR e Herculex, quando da utilização de sementes inteiras com níveis de contaminação de 0,2%. Foi possível, pela técnica de PCR convencional, detectar o evento TC1507...

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Section: Immunochromatographic Strip Test (Lateral Flow Membrane Strisupporting

confidence: 89%

Detection of transgenic events in maize using immunochromatographic strip test and conventional PCR

Cantelmo

Pinho

et al. 2013

Ciênc. agrotec.

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“…The size and color of AuNPs depend on the amount of sodium citrate that is used in the reduction of gold [17]. AuNPs with the diameter smaller than 15 nm were small for generating a strong color; however, nano-colloidal gold with a diameter of 20 nm was used as detector reagent.…”

Section: The Usage Methods Of Aunps In Lfamentioning

confidence: 99%

“…Kumar study. The better sensitivity was obtained with NC membrane of 5 μ pore size [17]. Another factor, which affects the sensitivity of LFA, is the amount of labeled-reaction antibody conjugates on the conjugate pad.…”

Section: Important Factors For Developing Lfamentioning

confidence: 99%

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Lateral flow assays: Principles, designs and labels

Bahadır

Sezgintürk

2016

TrAC Trends in Analytical Chemistry

500

315

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“…Nucleic acids and proteins are the usual target in such detection methods. Proteins are most widely used in the rapid tests, mainly due to the high reliability of the biochemical and protein-based detection methods [9], that include the enzyme-linked immunosorbent assay (ELISA) [10][11][12], fluorescence-linked immunosorbent assay (FLISA) [13], a bioassay using insect larvae [14] and lateral flow immunoassay [15]. Despite of success of those techniques they are laborious and it is required a specialized personnel.…”

Section: Introductionmentioning

confidence: 99%

Impedimetric immunosensors for the detection of Cry1Ab protein from genetically modified maize seeds

Freitas

Correr

Cancino-Bernardi

et al. 2016

Sensors and Actuators B: Chemical

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Development of nanocolloidal gold based immunochromatographic assay for rapid detection of transgenic vegetative insecticidal protein in genetically modified crops

Cited by 36 publications

References 17 publications

Detection of transgenic events in maize using immunochromatographic strip test and conventional PCR

Detection of transgenic events in maize using immunochromatographic strip test and conventional PCR

Lateral flow assays: Principles, designs and labels

Impedimetric immunosensors for the detection of Cry1Ab protein from genetically modified maize seeds

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