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PubMed1999

Rapid screening for diuretic doping agents in urine by C60-assisted laser-desorption-ionization-time-of-flight mass spectrometry.

This study describes a matrix-assisted laser-desorption-ionization (MALDI) mass spectrometry for rapid screening of 12 diuretics in spiked urine. C60 is used as the matrix for MALDI. Diuretics are directly analyzed by C60-MALDI without previous derivatization. The fact that most diuretic molecules contain sulfate groups accounts for why anions of the molecules can be easily desorbed and ionized in MALDI. Using C60 as the matrix is advantageous because of the low background in the low mass range on the negative MALDI mass spectrum. A clear mass window between m/z 200 and 600 in negative ion mode is also obtained. Only a minimum amount of the sample (< 1 microL) is necessary to perform the analysis. The detection limit of diuretics is approximately 0.1-1 microg/mL.

Benzothiadiazines↗
PubMed2002

Rapid screening of pKa values of pharmaceuticals by pressure-assisted capillary electrophoresis combined with short-end injection.

A method applying pressure-assisted capillary electrophoresis combined with short-end injection has been developed for the rapid screening of the pKa values of pharmaceuticals. The electrophoretic separation is performed on a short capillary length with short-end injection under an applied pressure, and the effective mobility is measured in a series of 10 different buffers with constant ionic strength (I = 0.05). The application of pressure not only reduces migration times, particularly in lower pH buffers, but also improves the repeatability of effective mobility measurements. The influence of pressure on the effective mobility was investigated at various pH values. It was observed for the first time that an increase in pressure resulted in a slight decrease in the effective mobility when the pH was above the pKa for acidic analytes, whereas an increased effective mobility with increasing pressures was observed when the pH was below the pKa. However, the observed effective mobility shift by the applied pressure did not significantly affect the determined pKa values. The determined pKa values were in good agreement with published data. Furthermore, a stacking condition was applied to increase the sensitivity, and a concentration down to 2 microM could readily be detected with UV detection using a 50 microm I.D. capillary. This technique is particularly suitable for measurement of pKa values for compounds with poor aqueous solubility. The method also omits the commonly used preconditioning steps with sodium hydroxide and water. The exclusion of excessive preconditioning steps and the use of pressure reduces the total cycling analysis time, and makes it possible to determine the pKa in less than 40 min per compound without loss of accuracy.

Electrolytes↗
PubMed1978

Determination of disaccharides in feces by permethylation and gas chromatography: rapid screening for carbohydrate intolerance in children.

Disaccharides can be identified and determined quantitatively in fecal specimens by permethylation of the components in crude supernatants followed by isothermal gas chromatographic separation on OV-17 columns. The method provides a rapid screening procedure for detecting patients suspected of having a carbohydrate intolerance secondary to intestinal disaccharidase deficiency.

Carbohydrate Metabolism, Inborn Errors↗
PubMed2005

Rapid screening of the entire mitochondrial DNA for low-level heteroplasmic mutations.

Alterations of the mitochondrial DNA (mtDNA) are implicated in various pathological conditions. In this study, we used denaturing high performance liquid chromatography (DHPLC) as a method to rapidly screen the entire mtDNA for mutations. Overlapping DNA fragments, amplified by one single cycling protocol from frozen pre-formulated PCR mixes, were subjected to DHPLC analysis. Single DHPLC injections of fragments yielded straightforward interpretation of results with a detection limit down to 1% mtDNA heteroplasmy. Furthermore, collection and re-amplification of low degree heteroduplex peak-fractions allowed sequence analysis of mtDNA mutations down to the detection limit of the DHPLC method. In order to demonstrate that the method has diagnostic value, we analyzed and confirmed known mtDNA mutations in patient samples.

Chromatography, High Pressure Liquid↗
PubMed1983

Micro-counterimmunoelectrophoresis: a rapid screening technique for trypanosomiasis.

A rapid and economical micro-counterimmunoelectrophoresis (MCIE) test, for antibody to trypanosome antigens, has been developed which may prove useful in trypanosomiasis surveillance. MCIE was more sensitive and more rapid than immunodiffusion (ID). Serum samples from trypanosome-infected rabbits and cattle (Nigerian and Gambian), from Kuwaiti dogs and camels, from Ivory Coast sleeping sickness patients, from Brazilian patients with Chagas's disease, plus mouse-anti-malaria sera have been tested by MCIE with Trypanosoma brucei, T. vivax, T. congolense and T. cruzi antigens. A few of the rabbit and the mice sera were also tested against Trichomonas vaginalis, Entamoeba histolytica, Plasmodium berghei or P. chabaudi antigens. Whenever possible the MCIE results were compared with parasitological or other serological tests (ID; indirect fluorescent antibody test, IFAT; enzyme-linked immunosorbent assay, ELISA; complement fixation test, CFT or indirect haemagglutination test, IHAT). The MCIE test was not species specific but the homologous species reactions were more often positive than heterologous ones. Homologous species reactions, with rabbit sera, detected antibodies as early as 10 days after infection and developed more precipitin lines as the infection progressed. Trypanosomal antigens and antisera cross-reacted with those of other protozoa (E. histolytica, Tr. vaginalis, P. chabaudi, P. berghei, and Babesia sp. and Leishmania sp.) but such reactions required an antigen concentration 4 to 16 times greater than that required for homologous trypanosome reactions (a factor which might be exploited for identification of different species of trypanosome).(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1975

Rapid screening method for aflatoxins and zearalenone in corn.

The methanol-water extraction system used in AOAC Method II for aflatoxins extracts both the aflatoxins and zearalenone from corn. Using this methanol-water extraction system as a base, a rapid screening procedure has been developed for these mycotoxins. The methanol-water extract is defatted with hexane and the pigments are precipitated with copper carbonate. The aflatoxins and zearalenone are subsequently extracted into chloroform and are then detected by half-plate TLC. An elapsed time of about 1 hr is required to analyze 1 sample. The sensitivity of the method is about 2 mu-g/kg for aflatoxin B-1 and 100 mu-g/kg for zearalenone.

Aflatoxins↗
PubMed1997

The DNA 'comet assay' as a rapid screening technique to control irradiated food.

The exposure of food to ionizing radiation is being progressively used in many countries to inactivate food pathogens, to eradicate pests, and to extend shelf-life, thereby contributing to a safer and more plentiful food supply. To ensure free consumer choice, irradiated food will be labelled as such, and to enforce labelling, analytical methods to detect the irradiation treatment in the food product itself are desirable. In particular, there is a need for simple and rapid screening methods for the control of irradiated food. The DNA comet assay offers great potential as a rapid tool to detect whether a wide variety of foodstuffs have been radiation processed. In order to simplify the test, the agarose single-layer set-up has been chosen, using a neutral protocol. Interlaboratory blind trials have been successfully carried out with a number of food products, both of animal and plant origin. This paper presents an overview of the hitherto obtained results and in addition the results of an intercomparison test with seeds, dried fruits and spices are described. In this intercomparison, an identification rate of 95% was achieved. Thus, using this novel technique, an effective screening of radiation-induced DNA fragmentation is obtained. Since other food treatments also may cause DNA fragmentation, samples with fragmented DNA suspected to have been irradiated should be analyzed by other validated methods for irradiated food, if such treatments which damage DNA cannot be excluded.

Animals↗
PubMed2002

Use of direct Gram stain of stomach biopsy as a rapid screening method for detection of Helicobacter pylori from peptic ulcer and gastritis patients.

Four hundred and thirty five stomach mucosal biopsies were taken from 145 consecutive patients (3 from each patient) during investigations for dyspepsia in three hospitals in Western Nigeria. The aim was to determine the best suited rapid screening method to aid fast diagnosis of ulcer/gastritis in this environment, using Gram stain, CLO test kit (urease production test) and culture methods. Eighty-nine (61.4%) biopsies were positive using Gram stain, 61 (42.1%) using CLO test kit and only 28 (19.3%) using culture. Based on the various limitations of CLO test kits and culture methods, Gram stain was adjudged the best suited rapid method. The clinical implication of this finding is discussed.

Bacteriological Techniques↗
PubMed1994

General high-performance liquid chromatographic procedures for the rapid screening of natural and synthetic corticosteroids.

A study of a multiple high-performance liquid chromatographic procedure for the separation of 30 different corticosteroids is described. Normal- and reversed-phase general methods operating in a wide polarity range have been developed for the rapid screening of multicomponent mixtures. The complementary application of both normal- and reversed-phase methods could permit clarification of uncertainty deriving from analyses performed by only one method.

Adrenal Cortex Hormones↗
PubMed1976

Rapid screening of lipid metabolism in monolayer cell cultures.

Monolayer cell cultures grown on coverslips in the presence of radioactive lipid precursors were embedded in silica gel layers for extraction and resolution of the labelled products directly by thin-layer chromatography. The method permits rapid screening of lipid metabolism in tissue cultures with a small number of cells.

Carbon Radioisotopes↗
PubMed2004

A method for rapidly screening functionality of actin mutants and tagged actins.

Recombinant production and biochemical analysis of actin mutants has been hampered by the fact that actin has an absolute requirement for the eukaryotic chaperone CCT to reach its native state. We therefore have developed a method to rapidly screen the folding capacity and functionality of actin variants, by combining in vitro expression of labelled actin with analysis on native gels, band shift assays or copolymerization tests. Additionally, we monitor, using immuno-fluorescence, incorporation of actin variants in cytoskeletal structures in transfected cells. We illustrate the method by two examples. In one we show that tagged versions of actin do not always behave native-like and in the other we study some of the molecular defects of three beta-actin mutants that have been associated with diseases.

Journal Article↗
PubMedDate not supplied

Cytologic detection of Cryptococcus neoformans in cerebrospinal fluid. Rapid screening methods.

Yeasts of Cryptococcus neoformans in cerebrospinal fluid (CSF) were studied using polarized light and fluorescence microscopy. Ultraviolet-induced fluorescence of wet-fixed, Papanicolaou-stained smears and polarization of air-dried, Diff-Quik or Leishman-stained preparations are complementary methods that allow for rapid screening for and immediate identification of Cryptococcus in cytologic preparations of CSF.

Birefringence↗
PubMed1993

Rapid screening test for enterotoxin-producing Bacillus cereus.

Culture supernatants of 30 enterotoxin-producing Bacillus cereus isolates produced a characteristic progressive destruction of McCoy cell monolayers. Enterotoxin-negative B. cereus and other group 1 Bacillus spp. caused no monolayer disruption. The McCoy cell tissue culture system appears to provide a rapid screening assay for detection of enterotoxin-producing B. cereus.

Animals↗
PubMed1980

Detection of human cytomegalovirus-specific IgG antibodies by a sensitive solid-phase radioimmunoassay and by a rapid-screening test.

A radioimmunoassay (RIA) for the detection of human IgG antibodies to cytomegalovirus (CMV) which utilizes extracts of CMV-infected cells as antigen is described. Sera of 66 healthy adults were titered by the RIA and by the complement fixation (CF) test. These same subjects were also used in the evaluation of a rapid screening radioimmunoassay (RSRIA) developed to detect IgG antibodies to CMV in small serum samples. The specificity of the RIA was evaluated by testing sera of seven CMV patients and ten patients with heterotypic titer rises to herpes simplex virus or varicella zoster virus (HSV or VZV). The RIA proved to be sensitive and specific. Possible applications of the RSRIA modification are discussed.

Antibodies, Viral↗
PubMedDate not supplied

Evaluation of a rapid screening assay for bacterial identification (Dot-ELISA) in fecal samples from children.

With the objective of standardizing a Dot Enzyme-Linked Immunosorbent Assay (Dot-ELISA) to detect antigens of fecal bacterial enteropathogens, 250 children, aged under 36 months and of both sexes, were studied; of which 162 had acute gastroenteritis. The efficacy of a rapid screening assay for bacterial enteropathogens (enteropathogenic Escherichia coli "EPEC", enteroinvasive Escherichia coli "EIEC", Salmonella spp. and Shigella spp.) was evaluated. The fecal samples were also submitted to a traditional method of stool culture for comparison. The concordance index between the two techniques, calculated using the Kappa (k) index for the above mentioned bacterial strains was 0.8859, 0.9055, 0.7932 and 0.7829 respectively. These values express an almost perfect degree of concordance for the first two and substantial concordance for the latter two, thus enabling this technique to be applied in the early diagnosis of diarrhea in infants. With a view to increasing the sensitivity and specificity of this immunological test, a study was made of the antigenic preparations obtained from two types of treatment: 1) deproteinization by heating; 2) precipitation and concentration of the lipopolysaccharide antigen (LPS) using an ethanol-acetone solution, which was then heated in the presence of sodium EDTA.

Child, Preschool↗
PubMed2001

Rapid screening for chiral separations by short-end injection capillary electrophoresis using highly sulfated cyclodextrins as chiral selectors.

Two series of amino acid derivatives and phenylamines were used to evaluate the potential of highly sulfated cyclodextrins (HS-CDs) for the screening for chiral separations by capillary electrophoresis (CE). HS-CDs showed to be very versatile and to exhibit very high enantioselectivity. The use of short-end injection allowed to reduce dramatically the analysis time. From the results obtained, a scheme for the rapid screening of enantiomeric molecules was developed and applied to various chiral drugs. Results are very satisfying as almost all compounds (62 out of 67) could be baseline-resolved. Usually, less than three experiments were necessary to obtain very good separation.

Acids↗
PubMed2001

The use of a Daphnia magna bioassay for rapid screening of acute intoxications with insecticides in dogs and cats.

Sudden death due to acute insecticide intoxications occurs frequently in dogs and cats. The absence of characteristic lesions at autopsy often renders post-mortem diagnosis dependent on the analysis of samples taken from the carcase at autopsy. In the present study, a bioassay utilizing Daphnia magna was proposed and tested as a rapid screening method for acute intoxications in dogs and cats. The bioassay was shown to be highly sensitive for detecting carbamate and organophosphate insecticides in the stomach contents. Generally. the mean survival time of the waterfleas in the control group was 5.17 h (SD = 1.24) and in the intoxicated group 1.32 h (SD = 1.49). during a 6 h observation period. If a cut-off is set at 4 h, this Daphnia bioassay gave 5.5% false negative results and 18.2% false positive results. using the results of toxicological analyses as a gold standard.

Animals↗
PubMedDate not supplied

Application of antibody adsorbed polyester cloth for the rapid screening of elution conditions for antigen immunopurification.

Antibody adsorbed onto polyester cloth did not desorb during a 20 hour agitated incubation in commonly used antigen-antibody dissociating agents. Antibody activity on the cloth was also unaffected by five 20 min cycles of agitation in these agents. Use of cloth segments coated with a model antibody, goat anti-rabbit IgG antibody, permitted rapid screening of elution conditions for the immunoaffinity purification of a model antigen, rabbit IgG. Thus, this simple test is useful for designing an optimal immunoaffinity chromatographic procedure.

Animals↗