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Selected AbstractsSimultaneous detection of genetically modified organisms by multiplex ligation-dependent genome amplification and capillary gel electrophoresis with laser-induced fluorescenceELECTROPHORESIS, Issue 13 2010Virginia García-Cañas Abstract In this work, an innovative method useful to simultaneously analyze multiple genetically modified organisms is described. The developed method consists in the combination of multiplex ligation-dependent genome dependent amplification (MLGA) with CGE and LIF detection using bare-fused silica capillaries. The MLGA process is based on oligonucleotide constructs, formed by a universal sequence (vector) and long specific oligonucleotides (selectors) that facilitate the circularization of specific DNA target regions. Subsequently, the circularized target sequences are simultaneously amplified with the same couple of primers and analyzed by CGE-LIF using a bare-fused silica capillary and a run electrolyte containing 2-hydroxyethyl cellulose acting as both sieving matrix and dynamic capillary coating. CGE-LIF is shown to be very useful and informative for optimizing MLGA parameters such as annealing temperature, number of ligation cycles, and selector probes concentration. We demonstrate the specificity of the method in detecting the presence of transgenic DNA in certified reference and raw commercial samples. The method developed is sensitive and allows the simultaneous detection in a single run of percentages of transgenic maize as low as 1% of GA21, 1% of MON863, and 1% of MON810 in maize samples with signal-to-noise ratios for the corresponding DNA peaks of 15, 12, and 26, respectively. These results demonstrate, to our knowledge for the first time, the great possibilities of MLGA techniques for genetically modified organisms analysis. [source] Application of extended inverse scatter correction to mid-infrared reflectance spectra of soilJOURNAL OF CHEMOMETRICS, Issue 5-7 2005Neal B. Gallagher Abstract Scattering artifacts adversely affect infrared reflectance measurements of powders and soils, and extended inverse scatter correction (EISC) is a flexible method useful for correcting these artifacts. EISC was used to correct mid-infrared reflectance spectra of two different soils coated with dibutyl phosphate and the results were examined using regression analysis. To obtain the correction, EISC fits a measured spectrum to a reference spectrum. However, if measured spectra contain features not included in the reference spectrum the fit can be biased resulting in poor correction. Weighted and robust least squares were used to account for these potential biases. Additionally, the present work demonstrates how analyte-free samples can be used to determine basis functions for an extended mixture model used in the correction. Corrected spectra resulted in partial least squares models that performed at least as well as 2nd derivative spectra and were more interpretable. Copyright © 2006 John Wiley & Sons, Ltd. [source] Item response theory: applications of modern test theory in medical educationMEDICAL EDUCATION, Issue 8 2003Steven M Downing Context Item response theory (IRT) measurement models are discussed in the context of their potential usefulness in various medical education settings such as assessment of achievement and evaluation of clinical performance. Purpose The purpose of this article is to compare and contrast IRT measurement with the more familiar classical measurement theory (CMT) and to explore the benefits of IRT applications in typical medical education settings. Summary CMT, the more common measurement model used in medical education, is straightforward and intuitive. Its limitation is that it is sample-dependent, in that all statistics are confounded with the particular sample of examinees who completed the assessment. Examinee scores from IRT are independent of the particular sample of test questions or assessment stimuli. Also, item characteristics, such as item difficulty, are independent of the particular sample of examinees. The IRT characteristic of invariance permits easy equating of examination scores, which places scores on a constant measurement scale and permits the legitimate comparison of student ability change over time. Three common IRT models and their statistical assumptions are discussed. IRT applications in computer-adaptive testing and as a method useful for adjusting rater error in clinical performance assessments are overviewed. Conclusions IRT measurement is a powerful tool used to solve a major problem of CMT, that is, the confounding of examinee ability with item characteristics. IRT measurement addresses important issues in medical education, such as eliminating rater error from performance assessments. [source] Flow cytometric detection of ,- D -glucuronidase gene in wild-type bacterial cells using in-situ PCRBIOTECHNOLOGY & BIOENGINEERING, Issue 2 2003Ramaiah Sachidanandham Abstract An in situ PCR-based flow cytometry method useful for monitoring the presence or absence of the ,- D -glucuronidase gene in Escherichia coli has been developed. A single-step fixation and permeabilization procedure, which maintained cell integrity at the elevated temperatures used during thermal cycling in the presence of PCR reagents, was demonstrated. We have chosen a shorter DNA sequence of length 147 bp for the PCR. Cells subjected to in situ PCR using fluorescein-12-dUTP as a label, showed the presence of uid both in epifluorescence microscopic examination and flow cytometric analysis. Multi-parametric analysis of flow cytometric profiles revealed that the efficiency of labeling was found to be high. The potential of in situ PCR for the detection of uid in intact coliform cells was then successfully tested with a fecal coliform isolated from the coastal waters of Singapore. © 2003 Wiley Periodicals, Inc. Biotechnol Bioeng 82: 127,133, 2003. [source] | ||||||||||