Verification of GMO analytical methods: DNA isolation and quantitative detection methods for Roundup Ready soy flour

2012 ◽  
Vol 29 ◽  
pp. S196-S197
Author(s):  
Remziye Yılmaz ◽  
Ceren Bayraç ◽  
Pelin Mutlu ◽  
Erdem Boy ◽  
Meral Yücel
Keyword(s):  
Analytical Methods ◽  
Dna Isolation ◽  
Soy Flour ◽  
Detection Methods ◽  
Quantitative Detection ◽  
Roundup Ready

scholarly journals Recovery of Mycobacteria from Heavily Contaminated Environmental Matrices

2021 ◽  
Vol 9 (10) ◽  
pp. 2178
Author(s):  
Vit Ulmann ◽  
Helena Modrá ◽  
Vladimir Babak ◽  
Ross Tim Weston ◽  
Ivo Pavlik
Keyword(s):  
Dna Isolation ◽  
Animal Origin ◽  
Quantitative Detection ◽  
Contamination Rate ◽  
Propidium Monoazide ◽  
Environmental Matrices ◽  
Tetradecyltrimethylammonium Bromide ◽  
Faecal Samples ◽  
Universal Procedure ◽  
Epidemiology Studies

For epidemiology studies, a decontamination method using a solution containing 4.0% NaOH and 0.5% tetradecyltrimethylammonium bromide (TDAB) represents a relatively simple and universal procedure for processing heavily microbially contaminated matrices together with increase of mycobacteria yield and elimination of gross contamination. A contamination rate only averaging 7.3% (2.4% in Cluster S; 6.9%% in Cluster R and 12.6%% in Cluster E) was found in 787 examined environmental samples. Mycobacteria were cultured from 28.5% of 274 soil and water sediments samples (Cluster S), 60.2% of 251 samples of raw and processed peat and other horticultural substrates (Cluster R), and 29.4% of 262 faecal samples along with other samples of animal origin (Cluster E). A total of 38 species of slow and rapidly growing mycobacteria were isolated. M. avium ssp. hominissuis, M. fortuitum, and M. malmoense were the species most often isolated. The parameters for the quantitative detection of mycobacteria by PCR can be significantly refined by treating the sample suspension before DNA isolation with PMA (propidium monoazide) solution. This effectively eliminates DNA residue from both dead mycobacterial cells and potentially interfering DNA segments present from other microbial flora. In terms of human exposure risk assessment, the potential exposure to live non-tuberculous mycobacteria can be more accurately determined.

Laser-Generated Surface Acoustic Wave Technique for Crack Monitoring – A Review

2013 ◽  
Vol 7 (2) ◽  
pp. 211-220 ◽  
Author(s):  
Kun Chen ◽  
◽  
Xing Fu ◽  
Dante J. Dorantes-Gonzalez ◽  
Yanning Li ◽  
...  
Keyword(s):  
Acoustic Wave ◽  
Surface Acoustic Wave ◽  
Acoustic Waves ◽  
Numerical Models ◽  
Surface Acoustic Waves ◽  
Detection Methods ◽  
Quantitative Detection ◽  
Quantitative Monitoring ◽  
Crack Monitoring ◽  
Wave Technique

In this paper, the principle of surface acoustic wave techniques and their application to the monitoring of cracks are presented and compared to other classic non-destructive techniques. A practical classification of methods regarding the excitation and detection of surface acoustic waves is enumerated, among them, laser-generated surface acoustic wave technique is carefully analyzed as a prospective technique, and two important detection methods using piezoelectric and light deflection are described. Then, the strategies and variables used in crack monitoring based on laser-generated surface acoustic wave technique are reviewed. To achieve the goal of quantitative detection of cracks, most researchers use numerical models and experiments to characterize main crack features. Discussions and prospective approaches for further quantitative monitoring of cracks are provided.

Qualitative and Quantitative PCR-Based Detection Methods for Authorized Genetically Modified Cotton Events in India

2014 ◽  
Vol 97 (5) ◽  
pp. 1299-1309 ◽  
Author(s):  
Rashmi Chhabra ◽  
Gurinder Jit Randhawa ◽  
Rajesh K Bhoge ◽  
Monika Singh
Keyword(s):  
Genetically Modified ◽  
Cost Effective ◽  
Detection Methods ◽  
Quantitative Detection ◽  
Bt Cotton ◽  
Pcr Assay ◽  
Genetically Modified Cotton ◽  
Detection Assay ◽  
Endogenous Reference ◽  
The Cost

Abstract Qualitative diagnostics for all five commercialized genetically modified (GM) cotton events for insect resistance in India is being reported for the first time in this paper. The cost-effective and robust multiplex PCR (MPCR)-based detection assay, distinguishing the insect resistant transgenic Bt cotton events, viz., MON531, MON15985, Event 1, GFM-cry1A, and MLS-9124, has been developed. This decaplex PCR assay targets nine transgenic elements, viz., sequences of four transgenes, three transgene constructs, and two event-specific sequences along with one endogenous reference gene. The LOD of the qualitative MPCR assay was up to 0.1%. A quantitative detection method for four widely commercially cultivated GM cotton events, namely, MON531, MON15985, Event 1, and GFM-cry1A, covering 99.5% of the total area under GM cultivation in the country, is also reported. A construct-specific real-time PCR assay has been developed for quantification of these GM cotton events with LOQ <0.05% and LOD <0.025%. The developed assays will be of great use to screen for the presence/absence of authorized GM cotton events in unknown samples and to check the authenticity of GM cotton seed samples.

scholarly journals Comparison of culture-based, vital stain and PMA-qPCR methods for the quantitative detection of viable hookworm ova

2017 ◽  
Vol 75 (11) ◽  
pp. 2615-2621 ◽  
Author(s):  
P. Gyawali ◽  
J. P. S. Sidhu ◽  
W. Ahmed ◽  
P. Jagals ◽  
S. Toze
Keyword(s):  
Environmental Samples ◽  
Quantitative Measurement ◽  
Quantitative Polymerase Chain Reaction ◽  
Detection Methods ◽  
Quantitative Detection ◽  
Propidium Monoazide ◽  
Chain Reaction ◽  
Vital Stain ◽  
Polymerase Chain ◽  
Hookworm Ova

Accurate quantitative measurement of viable hookworm ova from environmental samples is the key to controlling hookworm re-infections in the endemic regions. In this study, the accuracy of three quantitative detection methods [culture-based, vital stain and propidium monoazide-quantitative polymerase chain reaction (PMA-qPCR)] was evaluated by enumerating 1,000 ± 50 Ancylostoma caninum ova in the laboratory. The culture-based method was able to quantify an average of 397 ± 59 viable hookworm ova. Similarly, vital stain and PMA-qPCR methods quantified 644 ± 87 and 587 ± 91 viable ova, respectively. The numbers of viable ova estimated by the culture-based method were significantly (P < 0.05) lower than vital stain and PMA-qPCR methods. Therefore, both PMA-qPCR and vital stain methods appear to be suitable for the quantitative detection of viable hookworm ova. However, PMA-qPCR would be preferable over the vital stain method in scenarios where ova speciation is needed.

scholarly journals Analysis of trichloroethene vapour in soil-gas samples using solid-sorbent tubes with gas chromatography/mass spectrometry

2017 ◽  
Vol 14 (8) ◽  
pp. 495 ◽  
Author(s):  
Candice M. Duncan ◽  
Jon Mainhagu ◽  
Dan Lin ◽  
Mark L. Brusseau
Keyword(s):  
Mass Spectrometry ◽  
Gas Chromatography ◽  
Analytical Methods ◽  
Sampling Method ◽  
Significant Risk ◽  
Ambient Air ◽  
Soil Gas ◽  
Quantitative Detection ◽  
Sample Collection ◽  
Solid Sorbent

Environmental contextChlorinated chemicals are priority contaminants that pose significant risk to human health, and require state-of-the-art sampling techniques for varying matrices. A soil-gas sampling method was developed for the quantification of vapours of trichloroethene, a major chlorinated contaminant, present just above the groundwater zone. The method addresses sampling times, volumes and low-level trichloroethene concentrations. AbstractA sampling method for determining vapour concentrations of chlorinated contaminants, specifically trichloroethene (TCE), present in the vadose zone has been developed, and was applied at the Tucson International Airport Area Superfund site. The method, modified from the National Institute for Occupational Safety and Health (NIOSH) Manual of Analytical Methods # 1022 for ambient-air sampling of TCE, is targeted to situations requiring cost-effective sample collection, particularly for cases in which concentrations are at or below maximum contaminant. In our method, TCE vapour is sampled using a solid-sorbent tube. Gas chromatography with mass spectrometry is used to confirm and quantify the presence of TCE. The results of laboratory tests demonstrate a maximum TCE vapour load of ~22 mg before breakthrough to the secondary sorbent-tube section, and an extraction efficiency of ~97%. The results of a performance comparison test conducted in the field show that concentrations obtained with the sorbent tube samplers (~5 μg/L) are similar to those obtained with the use of standard summa canisters (~3 μg/L). The quantitative detection limit for the new method was 0.03 μg/L under the operative conditions, a significant improvement on current analytical methods. The results indicate that use of the sorbent-tube method will be effective for vapour sample collection at sites contaminated with volatile organic compounds, particularly in characterising low concentrations for applications such as assessing groundwater contamination risk and the need for remedial action via soil vapour extraction or other methods.

Sensitive Detection of Soy (Glycine max) by Real-Time Polymerase Chain Reaction Targeting the Mitochondrial atpA Gene

2011 ◽  
Vol 94 (6) ◽  
pp. 1863-1873 ◽  
Author(s):  
Tobias Bauer ◽  
Katja Kirschbaum ◽  
Silvia Panter ◽  
Marion Kenk ◽  
Jörg Bergemann
Keyword(s):  
Mitochondrial Dna ◽  
Real Time ◽  
Nuclear Dna ◽  
Mitochondrial Gene ◽  
Soy Flour ◽  
Detection Methods ◽  
Minimal Amount ◽  
Food Matrix ◽  
Atpa Gene ◽  
Reporter Probe

Abstract Detection of trace amounts of allergens is essential for correct labeling of food products by the food industry. PCR-based detection methods currently used for this purpose are targeting sequences of DNA present in the cell nucleus. In addition to nuclear DNA, a substantial amount of mitochondrial DNA (mtDNA) copies are present in the cytoplasm of eukaryotic cells. The nuclear DNA usually consists of a set of DNA molecules present in two copies per cell, whereas mitochondrial DNA is present in a few hundred copies per cell. Thus, an increase in sensitivity can be expected when mtDNA is used as the target. In this study, we present a reporter probe-based real-time PCR method amplifying the mitochondrial gene of the alpha chain of adenosine triphosphate synthetase from soy. Increase in sensitivity was examined by determining the minimal amount of soy DNA detectable by mtDNA and nuclear DNA (nDNA) amplification. Additionally, the LOD of soy in a food matrix was determined for mtDNA amplification and compared to the LOD determined by nDNA amplification. As food matrix, a model spice spiked with soy flour was used. Sensitivity of PCR-based soy detection can be increased by using mtDNA as the target.

scholarly journals Chemiluminescence Immunoassay Based Serological Immunoassays for Detection of SARS-CoV-2 Neutralizing Antibodies in COVID-19 Convalescent Patients and Vaccinated Population

Viruses ◽  
2021 ◽  
Vol 13 (8) ◽  
pp. 1508
Author(s):  
Qiangling Yin ◽  
Yecheng Zhang ◽  
Lijun Lian ◽  
Yuanyuan Qu ◽  
Wei Wu ◽  
...  
Keyword(s):  
Neutralizing Antibodies ◽  
Neutralizing Antibody ◽  
Detection Methods ◽  
Quantitative Detection ◽  
Antibody Concentration ◽  
Inhibition Assay ◽  
Healthy Donors ◽  
Kappa Value ◽  
Human Sera ◽  
Highly Correlated

The development of rapid serological detection methods re urgently needed for determination of neutralizing antibodies in sera. In this study, four rapid methods (ACE2-RBD inhibition assay, S1-IgG detection, RBD-IgG detection, and N-IgG detection) were established and evaluated based on chemiluminescence technology. For the first time, a broadly neutralizing antibody with high affinity was used as a standard for the quantitative detection of SARS-CoV-2 specific neutralizing antibodies in human sera. Sera from COVID-19 convalescent patients (N = 119), vaccinated donors (N = 86), and healthy donors (N = 299) confirmed by microneutralization test (MNT) were used to evaluate the above methods. The result showed that the ACE2-RBD inhibition assay calculated with either ACE2-RBD binding inhibition percentage rate or ACE2-RBD inhibiting antibody concentration were strongly correlated with MNT (r ≥ 0.78, p < 0.0001) and also highly consistent with MNT (Kappa Value ≥ 0.94, p < 0.01). There was also a strong correlation between the two evaluation indices (r ≥ 0.99, p < 0.0001). Meanwhile, S1-IgG and RBD-IgG quantitative detection were also significantly correlated with MNT (r ≥ 0.73, p < 0.0001), and both methods were highly correlated with each other (r ≥ 0.95, p < 0.0001). However, the concentration of N-IgG antibodies showed a lower correlation with the MNT results (r < 0.49, p < 0.0001). The diagnostic assays presented here could be used for the evaluation of SARS-CoV-2 vaccine immunization effect and serological diagnosis of COVID-19 patients, and could also have guiding significance for establishing other rapid serological methods to surrogate neutralization tests for SARS-CoV-2.

scholarly journals Genetic aspects of HPV infection detection in tumor and adjacent tissues of patients with laryngeal squamous cell carcinoma

2016 ◽  
Vol 61 (6) ◽  
pp. 275-279
Author(s):  
O. Yu. Dvoryaninova ◽  
E. G. Nikitina ◽  
V. A. Bychkov ◽  
N. V. Litviakov
Keyword(s):  
Squamous Cell Carcinoma ◽  
Cell Carcinoma ◽  
Squamous Cell ◽  
Laryngeal Squamous Cell Carcinoma ◽  
Hpv Infection ◽  
Detection Methods ◽  
Quantitative Detection ◽  
Tissue Samples ◽  
Hpv Dna ◽  
Advantages And Disadvantages

The article describes methods for the human papillomavirus (HPV) detection in tumor and adjacent (morphologically intact) tissues of patients with laryngeal squamous cell carcinoma (LSSC) in terms of viral pathogenesis. Comparative evaluation of the principles and techniques for HPV detection was performed. Advantages and disadvantages of the HPV detection methods are described. Approaches for DNA and HPV oncoproteins E6-E7 identification are substantiated. The results of our research into the qualitative and quantitative detection of HPV in the tumor and adjacent tissues of patients with Lssc are described. The research was conducted using commercial test systems Amplisens HPV HR screen-titre-FL and Amplisens HPV HR genotype-FL. Based on these results we developed the algorithm of HPV detection in samples of tumor tissue of patients with Lssc. The need for typing HPV-positive tissue samples with low concentration of HPV DNA was discussed.

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