Sensitive and highly specific quantitative real-time PCR and ELISA for recording a potential transfer of novel DNA and Cry1Ab protein from feed into bovine milk

  title={Sensitive and highly specific quantitative real-time PCR and ELISA for recording a potential transfer of novel DNA and Cry1Ab protein from feed into bovine milk},
  author={Patrick Guertler and Vijay Paul and Christiane Albrecht and Heinrich H. D. Meyer},
  journal={Analytical and Bioanalytical Chemistry},
AbstractTo address food safety concerns of the public regarding the potential transfer of recombinant DNA (cry1Ab) and protein (Cry1Ab) into the milk of cows fed genetically modified maize (MON810), a highly specific and sensitive quantitative real-time PCR (qPCR) and an ELISA were developed for monitoring suspicious presence of novel DNA and Cry1Ab protein in bovine milk. The developed assays were validated according to the assay validation criteria specified in the European Commission… 

A real-time immuno-PCR assay for the detection of transgenic Cry1Ab protein

  • R. Kumar
  • Biology
    European Food Research and Technology
  • 2011
The real-time immuno-PCR (IPCR) assay was developed for the detection and quantification of Cry1Ab protein and showed high sensitivity with minimum detection limit and found to be 10 times more sensitive than sandwich ELISA.

Development of ELISA for the detection of transgenic vegetative insecticidal protein in GM crops/produce.

  • R. Kumar
  • Biology
    Food additives & contaminants. Part A, Chemistry, analysis, control, exposure & risk assessment
  • 2012
The ELISA assay developed to support the labelling of vegetative insecticidal protein (Vip3A)-based GM produce was specific, sensitive and reproducible, which can be helpful to detect and track down the spread of unapproved and intentionally/unintentionally released GM produce harbouring Vip protein.

Establishment of a sandwich enzyme-linked immunosorbent assay for specific detection of Bacillus thuringiensis (Bt) Cry1Ab toxin utilizing a monoclonal antibody produced with a novel hapten designed with molecular model

A novel hapten polypeptide was synthesized and conjugated to keyhole limpet hemocyanin (KLH) and a monoclonal antibody named 2C12, showing high affinity to Cry1Ab and having no cross reaction with Cry1Ac, was produced, providing a new approach for the production of high specific antibody.

A novel sandwich ELISA system using rabbit monoclonal and polyclonal antibodies for rapid detection of Bt-Cry1Ac protein

The LOD and the linear detection range of the proposed ELISA are more satisfactory than other ELISA systems using an anti-Cry1Ac PcAb or two Cry1Ac-specific McAb for detection of the target protein and the research provides an effective instrument for detecting Cry1ac protein in GM crops and their derivatives.

Development of monoclonal antibody-based sensitive ELISA for the determination of Cry1Ie protein in transgenic plant

The results in this study suggested that the established ELISA method is effective for detecting Cry1Ie protein in transgenic plants.

Potential of cross-priming amplification and DNA-based lateral-flow strip biosensor for rapid on-site GMO screening

An on-site method for rapid detection of cauliflower mosaic virus promoter (CaMV 35S), a common element present in most GMO, using cross-priming amplification (CPA) technology, which has great commercial prospects for rapid GMO screening of high-consumption food and agriculture products.

Phage-Mediated Competitive Chemiluminescent Immunoassay for Detecting Cry1Ab Toxin by Using an Anti-Idiotypic Camel Nanobody.

It is demonstrated that the competitive format based on phage-displayed anti-idiotypic nanobodies can provide an alternative strategy for Cry toxin detection, with negligible cross-reactivity for other Cry toxins.

Long-term feeding of genetically modified maize (MON810) - Metabolism of recombinant DNA and the novel protein in the dairy cow

Only in feces of the transgenic fed group, the Cry1Ab protein was found, and this protein underlies a faster degradation than other proteins in feed, suggesting that it underlies the faster degradation of the cry1Ab DNA and the novel protein.



Sensitive PCR analysis of animal tissue samples for fragments of endogenous and transgenic plant DNA.

An optimized DNA extraction protocol for animal tissues coupled with sensitive PCR methods was used to determine whether trace levels of feed-derived DNA fragments, plant and/or transgenic, are

Fate of recombinant DNA and Cry1Ab protein after ingestion and dispersal of genetically modified maize in comparison to rapeseed by fallow deer (Dama dama)

After uptake of GM maize, neither cry1Ab-specific gene fragments nor Cry1Ab protein were detected in the GI tract of fallow deer, indicating complete digestion of the GM maize.

Detection of genetically modified DNA sequences in milk from the Italian market.

Degradation of transgenic Cry1Ab DNA and protein in Bt-176 maize during the ensiling process.

The ensiling process markedly decreases the presence of long functional cry1Ab gene fragments and full size Cry1Ab protein in maize silage.

Degradation and possible carry over of feed DNA monitored in pigs and poultry

Additional PCR examinations using unhatched chicken embryos provided the first indication that neither chloroplast nor maize genes are present endogenously within the wild-type poultry genome, therefore, a transient transfer of short forage DNA into most poultry organs can be suspected.

Degradation of Cry1Ab protein from genetically modified maize in the bovine gastrointestinal tract.

It was concluded that Cry1Ab protein is degraded during digestion in cattle after two independent sets of gastrointestinal samples revealed the apparent discrepancy between the results obtained by ELISA and immunoblotting.

Real-time quantitative PCR detection of genetically modified Maximizer maize and Roundup Ready soybean in some representative foods.

A fast and quantitative method was developed to detect transgenic "Maximizer" maize "event 176" (Novartis) and "Roundup Ready" soybean (Monsanto) in food by real-time quantitative PCR. The use of the

Detection of transgenic and endogenous plant DNA in rumen fluid, duodenal digesta, milk, blood, and feces of lactating dairy cows.

Fragments of the rubisco gene were detected in the majority of samples analyzed in both the liquid and solid phases of ruminal and duodenal digesta, milk, blood, and feces, but rarely in blood.