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Journal of Immunoassay & Immunochemistry

Shunsuke Fujii, Osamu Morinaga, Takuhiro Uto, Shuichi Nomura, Yukihiro Shoyama
Immunoassay systems using monoclonal antibodies (mAbs) are one of the most useful techniques in the analytical, biochemical, and clinical fields. In this study, a combination enzyme-linked immunosorbent assay (ELISA) using both anti-glycyrrhizin and anti-liquiritin mAbs (anti-GL/Liq mixture mAbs) was developed for quality control of licorice and its products. The combination ELISA demonstrated high sensitivity, reproducibility, and specificity for the total content of GL and Liq by a single assay. The developed ELISA was effective and useful as the first screening method in the selection of high-quality licorice from the Glycyrrhiza species and in confirming the quality of licorice-containing Kampo medicines...
November 28, 2016: Journal of Immunoassay & Immunochemistry
Susanne Pribbenow, Tulsidas G Shrivastav, Martin Dehnhard
Enzyme-immunoassays (EIA) that detect fecal testosterone metabolites (fTM) are powerful tools to monitor gonadal activity non-invasively. However, a challenge with testosterone EIAs might be their potential for cross-reactivities with structurally similar glucocorticoid metabolites. Therefore, we aimed to verify the capability of four different testosterone EIAs to monitor fTM without reflecting changes in adrenocortical activity in spotted hyenas by analyzing fecal samples following testosterone and ACTH challenge tests...
November 17, 2016: Journal of Immunoassay & Immunochemistry
Adelina Rosca, Gabriela Anton, Luminita Ene, Iulia Iancu, Aura Temereanca, Cristian L Achim, Simona M Ruta
This study aimed to investigate the influence of antiretroviral therapy on methylation markers, in a group of HIV infected, heavily treated patients. Immune and molecular methods were used to investigate potential changes in methylation profile in DNA isolated from peripheral blood mononuclear cells collected from antiretroviral-experienced HIV infected patients and healthy controls. The percentage of 5-methylcytosine was inversely correlated with proviral DNA and active replication while DNMT1 (p=0.01) and DNMT3A (p=0...
November 17, 2016: Journal of Immunoassay & Immunochemistry
Bharti Jain, J Kumarasamy, C Gholve, Savita Kulkarni, M G R Rajan
We describe the development and validation of multianalyte immunoassays (MAIA) for three analytes, viz., thyroxine (T4), thyroid stimulating hormone (TSH) and thyroglobulin (Tg) essential for assessment of thyroid function but having widely varying molecular weights. Using polycarbonate (PC) track-etched membranes (TEM) as an immobilization support and (125)I as the tracer, both competitive assay for T4 and non-competitive assay for TSH and Tg were performed on the same TEM. MAIA was found to be highly sensitive and precise with clinically useful working range and correlated very well with individual analyte immunoassays...
November 1, 2016: Journal of Immunoassay & Immunochemistry
Ana Rodríguez-Rodríguez, Juan José Egea-Guerrero, Ángel Vilches-Arenas, Manuel Jesús Quintanilla-Vázquez, Francisco Murillo-Cabezas, M Ángeles Muñoz-Sánchez
Changes in Urotensin-II (U-II) concentration, a potent vasoconstrictor peptide, have been detected in various pathologies, but it has been impossible to define a normality range. We aimed to analyze the concordance and interchangeability between two enzyme immunoassay methods developed by Phoenix Pharmaceuticals, Inc. to measure U-II plasma concentration in rats: ELISA and fluorescent EIA. Assays resulted positively correlated (r = 0.850; p < 0.01). There was a significant difference between assays values (p < 0...
November 1, 2016: Journal of Immunoassay & Immunochemistry
Presentini Rivo
A new method has been developed to prepare horseradish peroxidase (HRP) conjugates, using bis(sulfosuccinimidyl)suberate (BS3) as cross-linking reagent in a two-step procedure. The enzyme is reacted with BS3, introducing active ester molecules into the enzyme itself, and it is then used directly to label amino-containing compounds without further treatment. The proteins involved in the conjugation undergo minimal modifications. The reaction conditions are evaluated, as well as studies on the conservation of the biological activities of the conjugated proteins and the stability of the conjugates in time...
October 31, 2016: Journal of Immunoassay & Immunochemistry
Maysam Mard-Soltani, Mohamad Javad Rasaee, AbdolKarim Sheikhi, Mehdi Hedayati
Designing novel antigens to rise specific antibodies for Thyroid Stimulating Hormone (TSH) detection is of great significance. A novel fusion protein consisting of the C termini sequence of TSH beta subunit and a fusion sequence was designed and produced for rabbit immunization. Thereafter, the produced antibodies were purified and characterized for TSH detection. Our results indicate that the produced antibody is capable of sensitive and specific detection of TSH with low cross reactivity. This study underscores the applicability of designed fusion protein for specific and sensitive polyclonal antibody production and the importance of selecting an amenable region of the TSH for immunization...
October 27, 2016: Journal of Immunoassay & Immunochemistry
Awoyelu E Hilda, Oloke Julius Kola, Oladipo E Kolawole
Hepatitis C virus is one of the emerging infectious diseases that can be transmitted through blood-to-blood contact. This study was carried out to determine the prevalence of anti-HCV antibodies among potential blood donors and pregnant women attending Bowen University Teaching Hospital (BUTH), Ogbomoso, Oyo State. Hospital based study was conducted from December 2014 to September 2015. The study group (N = 279) included potential blood donors and pregnant women. Data on socio-demographic characteristics and potential risk factors were collected using a structured questionnaire...
September 27, 2016: Journal of Immunoassay & Immunochemistry
Elisa María Castells Martínez, Ruben Del Valle, Ernesto Carlos González, Antonio Melchor, Pedro Lucio Pérez, Idania González, Adriana Carr, Kalet León
Human epidermal growth factor is a small peptide consisting of 53 amino acid residues, which stimulates cell proliferation and is associated with several human carcinomas. A simple sandwich-type ultramicroELISA assay (UMELISA), based on the advantages of high affinity reaction between streptavidin and biotin has been developed for the measurement of EGF in human serum samples. Strips coated with a high affinity monoclonal antibody directed against EGF are used as solid phase, to ensure the specificity of the assay...
September 16, 2016: Journal of Immunoassay & Immunochemistry
Jan E Binnie, Delwyn G Cooke, Leonard F Blackwell
An enzyme-linked immunosorbent assay (ELISA) for measurement of pregnanediol-3α-glucuronide (PdG) excretion rates in urine samples diluted to 150 mL/h before analysis is described. The sensitivity of the 9 optimized standard curves was 0.093 ± 0.070 μmol PdG/24 hr, with the multiple combined standard curves having a mean mid-point (EC50) of 6.88 μmol PdG/24 hr. The PdG threshold excretion rate of 7.0 μmol/24 hr, which is used as a marker for the end of fertility, was situated in the most accurate region of the standard curve...
September 16, 2016: Journal of Immunoassay & Immunochemistry
Kheir Eddine Kerboua, Fatma Haiba, Djamila Batouche
Treatment of atypical hemolytic uremic syndrome (aHUS) by the complement C5 inhibitor eculizumab (Soliris®) is highly effective but unfortunately, associated with an economic pressure on the health care systems even in high incomes countries. Despite spacing infusions has been proposed as the unique solution to minimize this economic impact, no reliable laboratory assays are available to tailor such therapy optimization. We aimed to propose and evaluate a complement composite marker for eculizumab efficacy monitoring in aHUS...
September 12, 2016: Journal of Immunoassay & Immunochemistry
David Zeman, Pavlína Kušnierová, Jana Bojková, František Všianský, Olga Zapletalová
IgG kappa and IgG lambda concentrations were quantified in 96 paired CSF and sera using Hevylite™ antibodies in an in-house developed sandwich ELISA method. In 56 of these samples, the results were compared with a qualitative isoelectric focusing/affinity-mediated immunoblotting assay for oligoclonal IgG kappa and IgG lambda. Normal IgG kappa/lambda ratio in the CSF was the same as in serum. In 19/33 patients with intrathecal oligoclonal IgG synthesis, skewed IgG kappa/lambda ratio was observed (increased in 16 and decreased in 3 cases)...
September 9, 2016: Journal of Immunoassay & Immunochemistry
Tsuyoshi Torii, Kentaku Kanemitsu, Akeo Hagiwara
Serum sialic acid (SA) levels are important for diagnosis, follow-up, and mechanistic analysis of malignant diseases. However, little is known about the levels of SA bound to serum IgM. Here, we isolated IgM from sera of healthy individuals and patients with cancer using DEAE chromatography and 8% polyethylene glycol precipitation. In this fraction, which contained partially purified IgM (recovery; 52%; purity: 25%), SA was quantified with fluorescence detection-HPLC (detection limit: 0.08 μM). SA levels in the IgM-enriched fraction was significantly higher in cancer patients (104 ± 27 µM) than in healthy individuals (81 ± 11 µM; p = 0...
September 6, 2016: Journal of Immunoassay & Immunochemistry
Tuba Ozgocer, Sedat Yildiz, Cihat Uçar
Non-invasive measurement of cortisol in saliva is of prime importance as it represents a bioavailable neuroendocrine marker for stress. Therefore, in this study, we developed an enzyme immune assay that was suitable for salivary cortisol measurements. For that purpose, rabbit polyclonal antibody was raised against cortisol-3-CMO:BSA conjugate. The test was based on competition of liquid phase cortisol with conjugated cortisol on the solid phase. Primary antibody was used to bind available sites on the conjugate, which was proportional to numbers of cortisol in liquid phase...
September 6, 2016: Journal of Immunoassay & Immunochemistry
Rafid Mustafa, Elliott Blumenthal
Experiments were conducted to examine the effects of turmeric on spleen cell proliferation. Cell suspensions of spleen cells from young and aged mice were treated with or without conconavalin A (Con-A) as a proliferation stimulant, and with and without turmeric (20 mg/ml) in different concentrations. Spleen cells from young mice that received turmeric showed significant increase in spleen cell proliferation (p<0.05), while spleen cells from aged mice that received turmeric showed no significant increase in T lymphocytes...
August 25, 2016: Journal of Immunoassay & Immunochemistry
Majid Asiabanha Rezaee, Mohammad Javad Rasaee, Javad Mohammadnejad
Human cardiac troponin I (cTni) is the gold marker for early diagnosis of myocardial infarction. In this regard, four immune-dominant epitopes of cTni were predicted and their 3D structures were determined. Thereafter, the competitive performance of the peptides was monitored with the developed polyclonal antibody-based indirect competitive ELISA; a half-maximal inhibitory concentration (IC50) of 0.49 (µg/mL) and detection limit of 0.037 (µg/mL) were achieved for recombinant cTni. The competitive ELISA determined sensitivity levels of 0...
August 11, 2016: Journal of Immunoassay & Immunochemistry
Opeyemi Mayowa Onilude, Sabri Mohd Yusoff, Benjamin Obukowho Emikpe, Polycarp Tanko, Salisi M Shahrom, Mohammed Effendy
Early and accurate diagnosis of Brucella melitensis is essential for the treatment and control of brucellosis both in animals and humans. The thrust for the development of a rapid diagnostic technique to overcome the limitations of conventional microbiological and serological tests brought about this investigation on the development and application of dot-ELISA for antigen and antibody detection in infected goats. Fifteen apparently healthy Boer aged 2-3 years which tested negative for brucellosis using PCR and ELISA, were grouped into A (10 goats infected intraocularly with 10(7) CFU of B...
August 5, 2016: Journal of Immunoassay & Immunochemistry
Mona Mlika, Nourane Kria, Emna Braham, Chokri Chebbi, Faouzi El Mezni
BACKGROUND: Multidisciplinary concertation is mandatory in order to assess interstitial pneumonias. The study of the bronchoalveolar lavage helps evoking a diagnosis according to the lavage profile. In lymphocytic alveolitis, immunocytochemistry or in flux cytometry are necessary in order to identify the different clusters of lymphocytes implicated. AIM: Our objective was to evaluate the profile of 31 lymphocytic alveolitis using 2 different techniques which are the immunocytochemistry and the in flow cytometry in order to evaluate the efficacy of each technique and to compare the different results to the final diagnoses...
July 29, 2016: Journal of Immunoassay & Immunochemistry
Jaime Pérez-Griera, Juan Carlos Andreu-Ballester, Alejandro Hueso Zarandieta, José García de la Asunción, Silvia Masquefa Bondia
Secretory immunoglobulin A (SIgA) in serum is possibly the best index of SIgA presence in mucosal secretions in digestive tract and the mirror of its immunologic barrier against many pathogenic aggressions. The measurement of salivary SigA alone may be affected by total salivary secretion and its final concentration in the gland lumen is probably not useful as a appropriate index of mucosal secretions in the digestive tract. The usefulness of the determination of SigA against various epitopes in serum from patients with various autoimmune disease has been demonstrated...
July 25, 2016: Journal of Immunoassay & Immunochemistry
Prashant Giribhattanavar, Kavitha Kumar, Siddarth Raajasekar, Nagarathna Chandrashekar, Shripad Patil
Mycobacterium tuberculosis (H37Ra) culture filtrate proteins (CFP) are explored as a diagnostic marker for tuberculous meningitis (TBM). Cerebrospinal fluid (CSF) samples from patients were categorized as confirmed (n=47), suspected (n=20) and non-TBM (n=25) cases. Immune response by western-blot revealed TBM CSF samples are having heterogeneous response to CFP. CFP ELISA was 92% sensitive and 38.30% specific. ODs of confirmed TBM and non-TBM cases were significantly different (P<0.0001) and also the suspected TBM and non-TBM cases (P=0...
July 25, 2016: Journal of Immunoassay & Immunochemistry
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