A Nylon Filter A-ELISA for Detecting Viruses in Water

1993 ◽  
Vol 27 (7-8) ◽  
pp. 135-141 ◽  
Author(s):  
Abidelfatah M. Nasser ◽  
Yehudit Elkana ◽  
Leon Goldstein
Keyword(s):  
Positive Result ◽  
Solid Phase ◽  
Enteric Viruses ◽  
High Specificity ◽  
Negative Results ◽  
Order Of Magnitude ◽  
Lower Background

This study was designed to develop a modification of A-ELISA performed in microtitre plates. Nylon filters have been utilized successfully as a solid phase for the performance of A-ELISA. The use of nylon filters resulted in lower background than nitro-cellulose and paper filters, indicating their suitability as a solid phase for developing A-ELISA. With enteric viruses, human rotaviruses and MS-2 coliphage, negative results were obtained, suggesting high specificity of the developed technique for poliovirus 1. The sensitivity of the developed A-ELISA has been shown to be at least one order of magnitude greater than ordinary ELISA. A positive result with the nylon A-ELISA can be obtained with samples containing 100-1000 pfu/ml of poliovirus. Up to date methods used for detecting viruses in water are elaborate, time consuming and costly. Applying the nylon A-ELISA may overcome some of these disadvantages.

Comparison of the Potency of 8-L-Arginine, 8-D-Arginine and 8-D-Homoarginine Vasopressin Analogs with Substituted Phenylalanine in Position 2

1994 ◽  
Vol 59 (6) ◽  
pp. 1439-1450 ◽  
Author(s):  
Miroslava Žertová ◽  
Jiřina Slaninová ◽  
Zdenko Procházka
Keyword(s):  
Amino Acid ◽  
Solid Phase Synthesis ◽  
Solid Phase ◽  
Basic Amino Acid ◽  
The Other ◽  
Side Chain ◽  
Inhibitory Potency ◽  
Phase Synthesis ◽  
Other Hand ◽  
Order Of Magnitude

An analysis of the uterotonic potencies of all analogs having substituted L- or D-tyrosine or -phenylalanine in position 2 and L-arginine, D-arginine or D-homoarginine in position 8 was made. The series of analogs already published was completed by the solid phase synthesis of ten new analogs having L- or D-Phe, L- or D-Phe(2-Et), L- or D-Phe(2,4,6-triMe) or D-Tyr(Me) in position 2 and either L- or D-arginine in position 8. All newly synthesized analogs were found to be uterotonic inhibitors. Deamination increases both the agonistic and antagonistic potency. In the case of phenylalanine analogs the change of configuration from L to D in position 2 enhances the uterotonic inhibition for more than 1 order of magnitude. The L to D change in position 8 enhances the inhibitory potency negligibly. Prolongation of the side chain of the D-basic amino acid in position 8 seems to decrease slightly the inhibitory potency if there is L-substituted amino acid in position 2. On the other hand there is a tendency to the increase of the inhibitory potency if there is D-substituted amino acid in position 2.

Trapping Effect on the Kinetic Critical Radius in Nucleation and Growth Processes

2014 ◽  
Vol 790-791 ◽  
pp. 97-102
Author(s):  
Zoltán Erdélyi ◽  
Zoltán Balogh ◽  
Gabor L. Katona ◽  
Dezső L. Beke
Keyword(s):  
Critical Nucleus ◽  
Solid Phase ◽  
Kinetic Monte Carlo ◽  
Mean Field ◽  
Transformation Process ◽  
Nucleus Size ◽  
Critical Nucleus Size ◽  
Acta Mater ◽  
Order Of Magnitude ◽  
The Difference

The critical nucleus size—above which nuclei grow, below dissolve—during diffusion controlled nucleation in binary solid-solid phase transformation process is calculated using kinetic Monte Carlo (KMC). If atomic jumps are slower in an A-rich nucleus than in the embedding B-rich matrix, the nucleus traps the A atoms approaching its surface. It doesn’t have enough time to eject A atoms before new ones arrive, even if it would be favourable thermodynamically. In this case the critical nucleus size can be even by an order of magnitude smaller than expected from equilibrium thermodynamics or without trapping. These results were published in [Z. Erdélyi et al., Acta Mater. 58 (2010) 5639]. In a recent paper M. Leitner [M. Leitner, Acta Mater. 60 (2012) 6709] has questioned our results based on the arguments that his simulations led to different results, but he could not point out the reason for the difference. In this paper we summarize our original results and on the basis of recent KMC and kinetic mean field (KMF) simulations we show that Leitner’s conclusions are not valid and we confirm again our original results.

scholarly journals Experiments to test the possibility of transmutation by electronic bombardment

1927 ◽  
Vol 114 (767) ◽  
pp. 289-292 ◽  
Keyword(s):  
Melting Point ◽  
Positive Result ◽  
Quartz Tube ◽  
Pure Preparation ◽  
Negative Results

In an earlier paper experiments were described which were carried out in order to test the production of gold from mercury, as reported by Miethe and Stammreich, and by Nagaoka. These experiments led to conclusively negative results, but it was considered desirable to repeat also the work of Smits and Karssen upon the conversion of lead into thallium and mercury. To this end it was decided to adapt the quartz tube apparatus with sealed in tungsten leads, already used in the case of mercury, and described and illustrated in the previous paper (interrupted arc method). While waiting for a specially pure preparation of lead, tin suggested itself as a suitable metal for further experimentation. Besides its low melting point, there is the further advantage that indium, the most probable product of any transmutation of tin, can be spectroscopically detected in exceedingly minute traces, whilst its occurrence is so limited that the danger of accidental contamination from the ordinary materials of the laboratory is much smaller than is the case with most elements. In this respect, a positive result with indium would be more convincing than the production of mercury reported by Smits.

Determination of cannabinoids in oral fluid and urine of “light cannabis” consumers: a pilot study

2018 ◽  
Vol 57 (2) ◽  
pp. 238-243 ◽  
Author(s):  
Roberta Pacifici ◽  
Simona Pichini ◽  
Manuela Pellegrini ◽  
Roberta Tittarelli ◽  
Flaminia Pantano ◽  
...  
Keyword(s):  
Pilot Study ◽  
Oral Fluid ◽  
Urine Samples ◽  
Screening Tests ◽  
Gas Chromatography Mass Spectrometry ◽  
Negative Results ◽  
A Value ◽  
Order Of Magnitude ◽  
Confirmation Analysis

Abstract Background In those countries where cannabis use is still illegal, some manufacturers started producing and selling “light cannabis”: dried flowering tops containing the psychoactive principle Δ-9-tetrahydrocannabinol (THC) at concentrations lower than 0.2% together with variable concentration of cannabidiol (CBD). We here report a pilot study on the determination of cannabinoids in the oral fluid and urine of six individuals after smoking 1 g of “light cannabis”. Methods On site screening for oral fluid samples was performed, as a laboratory immunoassay test for urine samples. A validated gas chromatography-mass spectrometry (GC-MS) method was then applied to quantify THC and CBD, independently from results of screening tests. Results On site screening for oral fluid samples, with a THC cut-off of 25 ng/mL gave negative results for all the individuals at different times after smoking. Similarly, negative results for urine samples screening from all the individuals were obtained. Confirmation analyses showed that oral fluid THC was in the concentration range from 2.5 to 21.5 ng/mL in the first 30 min after smoking and then values slowly decreased. CBD values were usually one order of magnitude higher than those of THC. THC-COOH, the principal urinary THC metabolite, presented the maximum urinary value of 1.8 ng/mL, while urinary CBD had a value of 15.1 ng/mL. Conclusions Consumers of a single 1 g dose of “light cannabis” did not result as positive in urine screening, assessing recent consumption, so that confirmation would not be required. Conversely, they might result as positive to oral fluid testing with some on-site kits, with THC cut-off lower than 25 ng/mL, at least in the first hour after smoking and hence confirmation analysis can be then required. No conclusions can be drawn of eventual chronic users.

scholarly journals False Negative Results in High Viremia Parvovirus B19-Samples Tested with Real-Time PCR

2010 ◽  
Vol 59 (2) ◽  
pp. 129-132 ◽  
Author(s):  
PIOTR GRABARCZYK ◽  
ALEKSANDRA KALIŃSKA ◽  
EWA SULKOWSKA ◽  
EWA BROJER
Keyword(s):  
Positive Result ◽  
Real Time ◽  
Real Time Pcr ◽  
Parvovirus B19 ◽  
False Negative ◽  
Immunocompromised Patients ◽  
Negative Results ◽  
False Negative Results ◽  
Parvovirus B 19 ◽  
The Way

Extremely high viremia is observed during some viruses infection, especialy in immunocompromised patients. False negative results of Parvovirus B 19 DNA tests performed with real-time PCR in high viremic samples are reported. The way of fluorescence diagrams analysis and algorithm of positive result confirmation to exclude such phenomenon are proposed.

Colorimetric solid-phase minisequencing assay illustrated by detection of alpha 1-antitrypsin Z mutation

1993 ◽  
Vol 39 (11) ◽  
pp. 2282-2287 ◽  
Author(s):  
L Harju ◽  
T Weber ◽  
L Alexandrova ◽  
M Lukin ◽  
M Ranki ◽  
...  
Keyword(s):  
Solid Phase ◽  
Colorimetric Assay ◽  
Point Mutations ◽  
High Specificity ◽  
Microtiter Plate ◽  
Test Results ◽  
Single Nucleotide ◽  
Nitrophenyl Phosphate ◽  
Simple Alternative ◽  
Alpha 1 Antitrypsin

Abstract In solid-phase minisequencing, a defined point mutation is detected in microtiter plate-immobilized DNA by a single nucleotide primer extension reaction. We have here developed the method into a colorimetric assay and applied it to the detection of the Z mutation of the alpha 1-antitrypsin gene. We used novel nucleoside triphosphates modified with dinitrophenyl (DNP) hapten, permitting detection by anti-DNP-alkaline phosphatase conjugate, with p-nitrophenyl phosphate as substrate. The Z mutation is detected in two reactions: DNP-labeled dCTP is incorporated when the template is normal, DNP-dUTP when the Z mutation is present. Both modified nucleotides were incorporated with high specificity and with an efficiency similar to that of unmodified nucleotides. The test results are measured by spectrophotometry, yielding quantitative absorbance values. Calculation of the ratio of C to U signal permitted unambiguous distinction of normal homozygous, ZZ homozygous, and ZM heterozygous genotypes. The colorimetric minisequencing assay is rapid, standardized, and automatable, and thus provides an accurate and simple alternative for the analysis of known point mutations.

An evaluation of four serum tests for pregnancy.

1983 ◽  
Vol 29 (3) ◽  
pp. 561-563 ◽  
Author(s):  
K W Ryder ◽  
R A Munsick ◽  
T O Oei ◽  
P C Young ◽  
H F Blackford
Keyword(s):  
Positive Result ◽  
Early Pregnancy ◽  
Beta Subunit ◽  
The Other ◽  
Analytical Sensitivity ◽  
Negative Results ◽  
Human Choriogonadotropin ◽  
Beta Hcg

Abstract We evaluated four pregnancy tests (Biocept-G, Beta-CG, Preg/Stat, and HCG-Beta Screen), using sera from 59 nonpregnant subjects and 77 patients with serum human choriogonadotropin beta-subunit (beta-hCG) concentrations ranging from 4 to 100 000 int. units/L. The results obtained for each test were compared with the results predicted on the basis of the sample's beta-hCG concentration and the beta-hCG concentration the manufacturer claimed necessary for a positive result (the test's analytical sensitivity). Biocept-G had the best sensitivity (100%), specificity (98.9%), and accuracy (99.2%). Beta-CG had the poorest sensitivity (86.4%), Preg/Stat the poorest specificity (87.5%), and accuracy (92.6%). We confirmed the manufacturer's claimed analytical sensitivity (200 int. units/L) for the Biocept-G procedure, but our calculated analytical sensitivity for the other tests was significantly different from that claimed by their manufacturers. Best results were obtained with Biocept-G, but with its analytical sensitivity of 200 int. units/L, samples from early pregnancy will give negative results. None of the pregnancy tests evaluated here will establish the presence or absence of early pregnancy with certainty.

Monoclonal antibodies used in solid-phase and liquid-phase assays, as exemplified by progesterone assay.

1987 ◽  
Vol 33 (8) ◽  
pp. 1331-1337 ◽  
Author(s):  
W Schramm ◽  
T Yang ◽  
A R Midgley
Keyword(s):  
Monoclonal Antibodies ◽  
Liquid Phase ◽  
Solid Phase ◽  
Protein A ◽  
High Specificity ◽  
Binding Properties ◽  
Affinity Constants ◽  
Hybridoma Cell Lines ◽  
One Year ◽  
Better Than

Abstract Two immunoglobulins secreted by hybridoma cell lines have been systematically investigated to determine if they could be used in solid-phase assays to give results comparable with those obtained by conventional liquid-phase radioimmunoassay (RIA). The antibodies, BQ.1 and BQ.2, bind with high specificity to the steroid hormone progesterone. The affinity constants, Ka, to 125I-labeled progesterone derivatives are 1.1 X 10(11) L/mol and 9.1 X 10(9) L/mol, respectively. Progesterone inhibited the binding of radioiodinated derivatives (amides of tyramine, histamine, and tyrosine methylester with 11 alpha-progesterone hemisuccinate) equally well. For solid-phase assays, we immobilized antibody BQ.1 via Protein A to different polystyrene surfaces (about 30 pg per tube at 50% inhibition of radiolabeled tracer). Under these conditions, the performance of this antibody for the quantification of progesterone was equivalent to that obtained in RIA. For the immobilized antibody BQ.2, only 1/10 of the amount used for optimal results in RIA was required in solid-phase assays. Binding of either antibody to the antigen was undiminished after several freezing-thawing cycles. When immobilized on solid matrices, both antibodies retained up to 95% of their binding properties for one year. Thus high-affinity, high-specificity monoclonal antibodies can be obtained for haptens and, when suitably immobilized, can be used in solid-phase assays with results equal to or better than those obtained with liquid RIA.

Preparation of Milk Samples for Immunoassay and Liquid Chromatographic Screening Using Matrix Solid-Phase Dispersion

1994 ◽  
Vol 77 (4) ◽  
pp. 848-854 ◽  
Author(s):  
Steven A Barker ◽  
Austin R Long
Keyword(s):  
Solid Phase ◽  
Screening Tests ◽  
Dairy Production ◽  
Milk Samples ◽  
Matrix Solid Phase Dispersion ◽  
Phase Dispersion ◽  
Negative Results ◽  
Microbial Inhibition ◽  
Receptor Assays ◽  
Liquid Chromatographic

Abstract The use of drugs to maintain the health and maximize the output of dairy cattle has made the monitoring of milk for such agents essential. Screening tests based on immunological, microbial inhibition, and bacterial receptor assays have been developed for the detection of violative levels of therapeutic substances. However, such assays are not infallible, and false positive or negative results can occur when contaminants bind receptors or compete for the binding of the target residues. Such effects may arise from dietary sources, diseases, or other variables. Thus, a violation by such a test is not definitive until further confirmation is obtained. Our laboratory has developed extraction procedures for several drugs used in dairy production. Our method uses matrix solid-phase dispersion (MSPD) to isolate drugs away from contaminants and to eliminate many possible interferences. MSPD can also be used to enhance the specificity of such assays by fractionating various classes of drugs that may cross-react. Similarly, such methods may be used for liquid chromatographic screening and confirmation of a suspect sample.

scholarly journals Rubella-specific IgM reactivity in sera from cases of infectious mononucleosis

1983 ◽  
Vol 90 (3) ◽  
pp. 407-413 ◽  
Author(s):  
P. Morgan-Capner ◽  
R. S. Tedder ◽  
J. E. Mace
Keyword(s):  
Infectious Mononucleosis ◽  
Rubella Virus ◽  
Solid Phase ◽  
Igm Antibody ◽  
Negative Results ◽  
Solid Phase Immunoassay ◽  
Antibody Capture ◽  
Positive Results

SUMMARYEight sera from 125 cases of infectious mononucleosis (IM) were reactive for rubella-specific IgM in an M-antibody capture radioimmunoassay. The reactivity of individual sera varied depending upon the source of the rubella antigen used in the assay. One serum gave strongly positive results with some rubella haemagglutinating antigens but negative results with others and may have contained an IgM antibody which was capable of distinguishing between strains of rubella virus.If the diagnosis of rubella is based solely on detection in solid-phase immunoassay of rubella-specific IgM, IM should be excluded.

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