Search results for: elisa kits
#28203534 2017/02/16 Save this To Up
Comparison of methanol and isopropanol as wash solvents for determination of hair cortisol concentration in grizzly bears and polar bears.Methodological differences among laboratories are recognized as significant sources of variation in quantification of hair cortisol concentration (HCC). An important step in processing hair, particularly when collected from wildlife, is the choice of solvent used to remove or "wash" external hair shaft cortisol prior to quantification of HCC. The present study systematically compared methanol and isopropanol as wash solvents for their efficiency at removing external cortisol without extracting internal hair shaft cortisol in samples collected from free-ranging grizzly bears and polar bears. Cortisol concentrations in solvents and hair were determined in each of one to eight washes of hair with each solvent independently. •There were no significant decreases in internal hair shaft cortisol among all eight washes for either solvent, although methanol removed detectable hair surface cortisol after one wash in grizzly bear hair whereas hair surface cortisol was detected in all eight isopropanol washes.•There were no significant differences in polar bear HCC washed one to eight times with either solvent, but grizzly bear HCC was significantly greater in hair washed with isopropanol compared to methanol.•There were significant differences in HCC quantified using different commercial ELISA kits commonly used for HCC determinations.
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#28189633 2017/02/12 Save this To Up
An effective and simplified DO-stat control strategy for production of rabies glycoprotein in Pichia pastoris.The glycoprotein (G-protein) of rabies virus is responsible for viral attachment to the host cell surface and induces virus neutralization antibodies. In the present study, the G-protein gene of rabies virus CVS strain was cloned, sequenced and expressed in the yeast, Pichia pastoris, as a secreted protein, using a simplified DO-stat control feeding strategy. This strategy involves the addition of methanol when the dissolved oxygen (DO) level rises above the setpoint avoiding methanol accumulation and oxygen limitation. The G-protein expression was evaluated by SDS-PAGE, ELISA, and western blot assays. Like native G-protein, the recombinant G-protein was found reactive when it was challenged against specific antibodies. The data indicate that the recombinant G-protein can be easily expressed and isolated, and may be useful as a safe source in the production of diagnostic kits and subunit vaccines to prevent rabies.
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#28189147 2017/02/12 Save this To Up
Optimization and Validation of ELISA for Pre-Clinical Trials of Influenza Vaccine.Testing of every new vaccine involves investigation of its immunogenicity, which is based on monitoring its ability to induce specific antibodies in animals. The fastest and most sensitive method used for this purpose is enzyme-linked immunosorbent assay (ELISA). However, commercial ELISA kits with whole influenza virus antigens are not available on the market, and it is therefore essential to establish an adequate assay for testing influenza virusspecific antibodies. We developed ELISA with whole influenza virus strains for the season 2011/2012 as antigens and validated it by checking its specificity, accuracy, linearity, range, precision, and sensitivity. The results show that we developed high-quality ELISA that can be used to test immunogenicity of newly produced seasonal or pandemic vaccines in mice. The pre-existence of validated ELISA enables shortening the time from the process of vaccine production to its use in patients, which is particularly important in the case of a pandemic.
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#28188944 2017/02/11 Save this To Up
Comparison of Fecal Calprotectin in Exclusively Breastfed and Formula or Mixed Fed Infants in the First Six Months of Life.We conducted this study to compare fecal calprotectin between exclusively breastfed and formula or mixed fed infants aged one month and six months. Sixty term infants were enrolled from the labor ward of Valiasr Hospital between Oct 2011 and July 2015 and their fecal calprotectin was checked by the ELISA method and Hycult biotech kits. The enrolled infants had a birth weight of 2500-4000 g and no perinatal insults or hospitalization. Stool sampling was done at 1±1 week and at 6n±1 months. The six-month infants had no recent disease, antibiotic use or vaccination. The mean fecal calprotectin was higher in exclusively breastfed infants at first and sixth months than formula and mixed fed infants (368.85±204.49 and 283.21±381.41 µg/g versus 152.59±139.13 and 113.62±92.75 µg/g respectively). (P=0.0001 and 0.018) Fecal calprotectin was higher in infants with GERD than healthy babies in the first and sixth months (P=0.0001 and 0.004). Based on the role of calprotectin in inflammation, its higher levels in exclusively breastfed infants is contrary to breast milk benefits and may be a sign of enhanced mucosal immune maturity in them.
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#28187301 2017/02/10 Save this To Up
The preparation of dual-functional hybrid nanoflower and its application in the ultrasensitive detection of disease-related biomarker.In this work, dual-functional streptavidin (SA)-horseradish peroxidase (HRP) hybrid nanoflowers, integrating the functions of biological recognition and signal amplification, were prepared through facile one-pot green synthesis method. The prepared SA-HRP-Cu3(PO4)2 hybrid nanoflowers loaded abundant HRP and simultaneously exhibited enhanced catalytic activity, stability, and durability compared with free enzyme, which fits greatly well with the requirement of signal tag for bioassay. Besides, due to the general SA-biotin linking interaction, the SA-HRP-Cu3(PO4)2 hybrid nanoflowers possess universal capture ability to the biotinylated antibody. Hence, combined with enzyme-linked immunosorbent assay (ELISA), the dual-functional hybrid nanoflowers were used to construct a colorimetric sensor for the ultrasensitive detection of alpha-fetoprotein (AFP). The detection limit is 78pg/mL, which is far superior to commercial ELISA kits. This presented approach holds great promise to develop on-demand hybrid system for a variety of applications ranging from biosensor and biomedicine to biocatalytic process.
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#28187244 2017/02/10 Save this To Up
Analytical performance of three diagnostic reagents for HBsAg on an automatic ELISA analyzer.We conducted performance tests of three HBsAg ELISA diagnostic reagents using an Addcare 600 (Yantai Addcare Bio-tech Limited Company) and studied the consistency between the qualitative results and chemiluminescent microparticle immunoassay (CMIA) results.
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#28180064 2017/02/09 Save this To Up
Dipeptidyl peptidase-4 inhibitor treatment induces a greater increase in plasma levels of bioactive GIP than GLP-1 in non-diabetic subjects.Glucose-dependent insulinotropic polypeptide (GIP) and glucagon-like peptide-1 (GLP-1) possess multiple bioactive isoforms that are rendered non-insulinotropic by the enzyme dipeptidyl peptidase-4 (DPP-4). Recently, some ELISA kits have been developed to specifically measure "active" GIP and GLP-1, but it is unclear if these kits can accurately quantify all bioactive forms. Therefore, it remains uncertain to what extent treatment with a DPP-4 inhibitor boosts levels of biologically active GIP and GLP-1. Thus, we evaluated our novel receptor-mediated incretin bioassays in comparison to commercially available ELISA kits using plasma samples from healthy subjects before and after DPP-4 inhibitor administration.
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#28165296 2017/02/06 Save this To Up
The activation of the kynurenine pathway in a rat model with renovascular hypertension.Hypertension is a serious condition that can lead to many health problems. The mechanisms underlying this process are still not fully understood. The kynurenine pathway may be involved in the occurrence and progression of hypertension. The purpose of this study was to examine the activity of peripheral kynurenine pathway in rats with renovascular hypertension in Goldblatt 2K1C model. Hypertension was induced in the experimental groups by constricting the renal artery of the left kidney of the rats. Determination of tryptophan (Trp) and kynurenine pathway metabolites was assessed by high-performance liquid chromatography in plasma and tissues obtained at 4, 8, and 16 weeks after the surgical intervention or sham surgery. Levels of Ang II were evaluated using commercial immuno-enzymatic ELISA kits. Surgical treatment led to increased values of mean blood pressure and systolic blood pressure, whereas Trp concentrations were decreased in experimental animals compared to appropriate controls. Simultaneously, the considerable increment of kynurenine pathway components and a significant increase in the activity of tryptophan 2,3-dioxygenase were observed in rats with developed hypertension in comparison with controls. There were no differences between Ang II levels in controls and experimental groups. The inverse relationship was between plasma Trp and both SBP and Ang II values, and Trp independently affected Ang II concentrations in hypertensive rats. In contrast, tryptophan 2,3-dioxygenase activity and plasma kynurenine metabolites positively correlated with blood pressure values as well as with Ang II levels in these animals. Moreover, kynurenine was independently connected with MBP. Renovascular hypertension influences kynurenine pathway and leads to an imbalance in Trp and its metabolite levels. Tryptophan 2,3-dioxygenase and part of the kynurenine metabolites in plasma and tissues positively correlated with blood pressure values and Ang II levels. Although the mechanisms underlying this phenomenon are unclear, our experiment showed a link between renovascular hypertension and activation of kynurenine pathway.
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#28164648 2017/02/06 Save this To Up
Validation of a New Panel of Automated Chemiluminescence Assays for Anticardiolipin Antibodies in the Screening for Antiphospholipid Syndrome.Antibodies anticardiolipin (aCL) and anti-β2-glycoprotein I (aβ2GPI) are two of three laboratory criteria of antiphospholipid syndrome (APS). All of assays of antiphospholipid antibodies (aPL), coagulation assays as well as ELISAs, show methodological shortcomings, that affect their sensitivity and specificity. Therefore, we decided to validate these parameters for a new chemiluminescent examination (CLIA).
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#28164613 2017/02/06 Save this To Up
High Concentrations of the Angiogenic Peptide VEGF-A in Seminal Fluid and its Association to Prostasomes.Angiogenesis is the formation of new blood vessels by capillary sprouting from pre-existing vessels. This process is associated with increased expression of angiogenic factors like vascular endothelial growth factor (VEGF). The VEGF family consists of five members denoted VEGF-A, B, C, D and placenta growth factor (PlGF). Prostasomes are exosome-like extracellular vesicles existing in seminal plasma. The present study aimed at investigating the possible relationship between VEGF-A in seminal fluid and blood plasma and the prostasomal association of VEGF-A.
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