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Immunodiagnostics

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https://www.readbyqxmd.com/read/28087312/immunisation-choice-of-host-adjuvants-and-boosting-schedules-with-emphasis-on-polyclonal-antibody-production
#1
REVIEW
Philippe Delahaut
Polyclonal antibodies are frequently used as immunodiagnostic tools in fundamental research. They are also used for routine diagnostic purposes in human and veterinary medicine and for quality control procedures in the food-processing industry. The antibody is a major component of the detection system. It binds with the molecule to be identified. This conjugate is subsequently revealed by means of binding the antibody with a radio-isotope, a fluorescent substance, an enzyme inducing a color change, or a biosensor based analytical system...
January 10, 2017: Methods: a Companion to Methods in Enzymology
https://www.readbyqxmd.com/read/28043501/comparative-proteomic-analysis-of-sequential-isolates-of-mycobacterium-tuberculosis-sensitive-and-resistant-beijing-type-from-a-patient-with-pulmonary-tuberculosis
#2
Amit Singh, Anil Kumar Gupta, Krishnamoorthy Gopinath, Divakar Sharma, Prashant Sharma, Deepa Bisht, Pawan Sharma, Sarman Singh
AIM & OBJECTIVE: In India, tuberculosis (TB) is a foremost health problem, and the emergence of multidrug-resistant (MDR) and extensively drug resistant (XDR) strains of Mycobacterium tuberculosis (M. tuberculosis) has further complicated the situation. Although various mechanisms have been proposed to elucidate the emergence of resistance, our knowledge remains insufficient. The formation of a very complex network and drugs of proteins are countered by their efflux/modification or target over-expression/modification...
December 2016: International Journal of Mycobacteriology
https://www.readbyqxmd.com/read/28043495/development-of-rapid-immuno-diagnostic-test-for-the-early-detection-of-tuberculosis
#3
Anil Kumar Gupta, Saumya Srivastava, Amit Singh, Sarman Singh
AIMS AND OBJECTIVE: To develop novel immunodiagnostic test for tuberculosis. METHODS: The selected novel proteins named as Rv2145c (SS1), Rv1437 (SS2), Rv1827 (SS3), and Rv2970c (SS4) were cloned, expressed and purified under specific conditions. Additionally, monoclonal antibodies (mAbs) were developed using these recombinant antigens via hybridoma technology. Hybridoma clones were screened and positive clones were selected for further experiment. The mAbs were purified from cell culture supernatant using protein A/G column chromatography...
December 2016: International Journal of Mycobacteriology
https://www.readbyqxmd.com/read/28043327/acetylcholinesterases-from-entomopathogenic-nematode-heterorhabditid-bacteriophora-susceptibility-to-insecticides-and-immunological-characteristics
#4
Magda A Mohamed, El-Sayed M E Mahdy, Abd-El-Hady M Ghazy, Nihal M Ibrahim, Hatem A El-Mezayen, Manal M E Ghanem
Acetylcholinesterases (AChEs) from the infective juveniles (IJs) of entomopathogenic nematode (EPN) have been investigated with respect to their susceptibility to insecticides and immunological characteristics, aiming at nominating the most compatible insecticide(s) to be used in conjunction with the most insecticide-tolerant EPN strain before incorporation in integrated pest management (IPM) programs. The inhibition kinetics of two purified AChE isoenzymes, AChEAII and AChEBI isolated from Heterorhabditid bacteriophora EM2 strain, by different insecticides revealed that the insensitivity to inhibition by such insecticides could be arranged in a descending order as; methomyl>carbofuran>acetamiprid>oxamyl>malathion...
January 2017: Pesticide Biochemistry and Physiology
https://www.readbyqxmd.com/read/28012899/corning-hyperflask-%C3%A2-for-viral-amplification-and-production-of-diagnostic-reagents
#5
Brian J Kearney, Matthew A Voorhees, Priscilla L Williams, Scott P Olschner, Cynthia A Rossi, Randal J Schoepp
Viral preparations are essential components in diagnostic research and development. The production of large quantities of virus traditionally is done by infecting numerous tissue culture flasks or roller bottles, which require large incubators and/or roller bottle racks. The Corning HYPERFlask(®) is a multilayer flask that uses a gas permeable film to provide gas exchange between the cells and culture medium and the atmospheric environment. This study evaluated the suitability of the HYPERFlask for production of Lassa, Ebola, Bundibugyo, Reston, and Marburg viruses and compared it to more traditional methods using tissue culture flasks and roller bottles...
December 21, 2016: Journal of Virological Methods
https://www.readbyqxmd.com/read/27936034/flow-cytometry-as-a-tool-for-quality-control-of-fluorescent-conjugates-used-in-immunoassays
#6
Marta de Almeida Santiago, Bruna de Paula Fonseca E Fonseca, Christiane de Fátima da Silva Marques, Edimilson Domingos da Silva, Alvaro Luiz Bertho, Ana Cristina Martins de Almeida Nogueira
The use of antibodies in immunodiagnostic kits generally implies the conjugation of these proteins with other molecules such as chromophores or fluorochromes. The development of more sensitive quality control procedures than spectrophotometry is essential to assure the use of better fluorescent conjugates since the fluorescent conjugates are critical reagents for a variety of immunodiagnostic kits. In this article, we demonstrate a new flow cytometric protocol to evaluate conjugates by molecules of equivalent soluble fluorochromes (MESF) and by traditional flow cytometric analysis...
2016: PloS One
https://www.readbyqxmd.com/read/27894367/potential-immunological-markers-for-diagnosis-of-human-strongyloidiasis-using-heterologous-antigens
#7
M A Corral, F M Paula, D M C L Meisel, V L P Castilho, E M N Gonçalves, D Levy, S P Bydlowski, P P Chieffi, W Castro-Borges, R C B Gryschek
Strongyloides venezuelensis is a parasitic nematode of rodents that is frequently used to obtain heterologous antigens for immunological diagnosis of human strongyloidiasis. The aim of this study was to identify antigens from filariform larvae of S. venezuelensis for immunodiagnosis of human strongyloidiasis. Soluble and membrane fractions from filariform larvae of S. venezuelensis were obtained in phosphate saline (SS and SM) and in Tris-HCl buffer (TS and TM), and were analysed by Western blotting. Different antigenic components were recognized by IgG antibodies from the sera of strongyloidiasis patients...
November 29, 2016: Parasitology
https://www.readbyqxmd.com/read/27881117/novel-prokaryotic-expression-of-thioredoxin-fused-insulinoma-associated-protein-tyrosine-phosphatase-2-ia-2-its-characterization-and-immunodiagnostic-application
#8
Luciano Lucas Guerra, Natalia Inés Faccinetti, Aldana Trabucchi, Bruno David Rovitto, Adriana Victoria Sabljic, Edgardo Poskus, Ruben Francisco Iacono, Silvina Noemí Valdez
BACKGROUND: The insulinoma associated protein tyrosine phosphatase 2 (IA-2) is one of the immunodominant autoantigens involved in the autoimmune attack to the beta-cell in Type 1 Diabetes Mellitus. In this work we have developed a complete and original process for the production and recovery of the properly folded intracellular domain of IA-2 fused to thioredoxin (TrxIA-2ic) in Escherichia coli GI698 and GI724 strains. We have also carried out the biochemical and immunochemical characterization of TrxIA-2icand design variants of non-radiometric immunoassays for the efficient detection of IA-2 autoantibodies (IA-2A)...
November 24, 2016: BMC Biotechnology
https://www.readbyqxmd.com/read/27876681/site-selective-orientated-immobilization-of-antibodies-and-conjugates-for-immunodiagnostics-development
#9
REVIEW
Min Shen, James F Rusling, Chandra K Dixit
Immobilized antibody systems are the key to develop efficient diagnostics and separations tools. In the last decade, developments in the field of biomolecular engineering and crosslinker chemistry have greatly influenced the development of this field. With all these new approaches at our disposal, several new immobilization methods have been created to address the main challenges associated with immobilized antibodies. Few of these challenges that we have discussed in this review are mainly associated to the site-specific immobilization, appropriate orientation, and activity retention...
November 19, 2016: Methods: a Companion to Methods in Enzymology
https://www.readbyqxmd.com/read/27853117/assessing-the-archaeoparasitological-potential-of-quids-as-a-source-material-for-immunodiagnostic-analyses
#10
Johnica J Morrow, Karl J Reinhard
In the present study, quids from La Cueva de los Muertos Chiquitos (CMC) were subjected to ELISA tests for 2 protozoan parasites, Toxoplasma gondii (n=45) and Trypanosoma cruzi (n=43). The people who occupied CMC, the Loma San Gabriel, lived throughout much of present-day Durango and Zacatecas in Mexico. The known pathoecology of these people puts them into at-risk categories for the transmission of T. gondii and T. cruzi. Human antibodies created in response to these 2 parasites can be detected in modern saliva using ELISA kits intended for use with human serum...
October 2016: Korean Journal of Parasitology
https://www.readbyqxmd.com/read/27787831/in-silico-prediction-of-linear-b-cell-epitopes-on-proteins
#11
Yasser El-Manzalawy, Drena Dobbs, Vasant G Honavar
Antibody-protein interactions play a critical role in the humoral immune response. B-cells secrete antibodies, which bind antigens (e.g., cell surface proteins of pathogens). The specific parts of antigens that are recognized by antibodies are called B-cell epitopes. These epitopes can be linear, corresponding to a contiguous amino acid sequence fragment of an antigen, or conformational, in which residues critical for recognition may not be contiguous in the primary sequence, but are in close proximity within the folded protein 3D structure...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/27748211/usefulness-of-gel-filtration-fraction-as-potential-biomarker-for-neurocysticercosis-in-serum-towards-a-new-diagnostic-tool
#12
D S Nunes, H T Gonzaga, V S Ribeiro, J P Cunha-Júnior, J M Costa-Cruz
There is an increasing interest in improving neurocysticercosis (NCC) diagnosis through the search of new and alternative antigenic sources, as those obtained from heterologous antigens. The aim of this study was to obtain potential biomarkers for NCC diagnosis after gel filtration chromatography [gel filtration fraction (GFF)] from the total saline extract (SE) from Taenia saginata metacestodes, followed by protein identification and application in immunodiagnostic. SE and GFF proteic profiles were characterized in gel electrophoresis, and diagnostic performance was verified by testing 160 serum samples through enzyme-linked immunosorbent assay and immunoblotting...
October 17, 2016: Parasitology
https://www.readbyqxmd.com/read/27746871/production-of-specific-igy-antibody-to-the-recombinant-fanc-protein-produced-in-escherichia-coli
#13
Khadijeh Nasiri, Saeed Zibaee, Mohammadreza Nassiri, Mojtaba Tahmoorespur, Alireza Haghparast
OBJECTIVES: Enterotoxigenic Escherichia coli (ETEC) strains are one of the primary causes of diarrhea in newborn calves and in humans, pigs, and sheep. IgY technology has been identified as a promising alternative to generating a mass amount of specific antibody for use in immunotherapy and immunodiagnostics. The purpose of this study was to produce specific antibody by egg yolk antibody (IgY) to recombinant FanC protein from ETEC. MATERIALS AND METHODS: FanC (K99) gene was amplified from ETEC by specific primers and polymerase chain reaction...
August 2016: Iranian Journal of Basic Medical Sciences
https://www.readbyqxmd.com/read/27733801/histopathological-and-immunohistochemical-approaches-for-the-diagnosis-of-pasteurellosis-in-swine-population-of-punjab
#14
Payal Bhat, Nittin Dev Singh, Geeta Devi Leishangthem, Amninder Kaur, Vishal Mahajan, Harmanjit Singh Banga, Rajinder Singh Brar
AIM: Infectious porcine bronchopneumonia, caused by Pasteurella multocida, is a widespread disease of major economic significance. Thus, the aim of the present study was to diagnose swine Pasteurellosis using gross, histopathological, and immunopathological approaches in the swine population of Punjab and to compare the efficacy of immunohistochemical (IHC) techniques with conventional diagnostic techniques. MATERIALS AND METHODS: A total of 71 adult swine lung samples showing gross pneumonic changes were collected along with the associated lymph nodes to carry out the study...
September 2016: Veterinary World
https://www.readbyqxmd.com/read/27730559/purification-of-antibodies-using-affinity-chromatography
#15
Elaine Darcy, Paul Leonard, Jenny Fitzgerald, Martin Danaher, Hui Ma, Richard O'Kennedy
Affinity chromatography permits the isolation of a target analyte from a complex mixture and can be utilized to purify proteins, carbohydrates, drugs, haptens, or any analyte of interest once an affinity pair is available. It involves the exploitation of specific interactions between a binding affinity pair, such as those between an antibody and its associated antigen, or between any ligand and its associated binding receptor/protein. With the discovery of protein A in 1970, and, subsequently protein G and L, immuno-affinity chromatography has grown in popularity and is now the standard methodology for the purification of antibodies which may be implemented for a selection of different applications such as immunodiagnostics...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/27726808/mycobacteria
#16
Patricia J Simner, Gail L Woods, Nancy L Wengenack
The immunocompromised host is at increased risk of Mycobacterium tuberculosis complex and nontuberculous mycobacteria infection. Although Mycobacterium tuberculosis complex is a significant mycobacterial pathogen, nontuberculous mycobacteria causes substantial disease in those with suppressed immune responses. Mycobacterial infections can cause significant morbidity and mortality in this patient population, and rapid identification and susceptibility testing of the mycobacterial species is paramount to patient management and outcomes...
August 2016: Microbiology Spectrum
https://www.readbyqxmd.com/read/27706156/validation-of-a-rapid-rabies-diagnostic-tool-for-field-surveillance-in-developing-countries
#17
Monique Léchenne, Kemdongarti Naïssengar, Anthony Lepelletier, Idriss Oumar Alfaroukh, Hervé Bourhy, Jakob Zinsstag, Laurent Dacheux
BACKGROUND: One root cause of the neglect of rabies is the lack of adequate diagnostic tests in the context of low income countries. A rapid, performance friendly and low cost method to detect rabies virus (RABV) in brain samples will contribute positively to surveillance and consequently to accurate data reporting, which is presently missing in the majority of rabies endemic countries. METHODOLOGY/PRINCIPAL FINDINGS: We evaluated a rapid immunodiagnostic test (RIDT) in comparison with the standard fluorescent antibody test (FAT) and confirmed the detection of the viral RNA by real time reverse transcription polymerase chain reaction (RT-qPCR)...
October 2016: PLoS Neglected Tropical Diseases
https://www.readbyqxmd.com/read/27685323/antibody-labeling-with-fluorescent-dyes-using-magnetic-protein-a-and-protein-g-beads
#18
Nidhi Nath, Becky Godat, Marjeta Urh
Antibodies labeled with small molecules like fluorescent dyes, cytotoxic drugs, and radioactive tracers are essential tools in biomedical research, immunodiagnostics and more recently as therapeutic agents. Traditional methods for labeling antibodies with small molecules require purified antibodies at relatively high concentration, involve multiple dialysis steps and have limited throughput. However, several applications, including the field of Antibody Drug Conjugates (ADCs), will benefit from new methods that will allow labeling of antibodies directly from cell media...
2016: Journal of Visualized Experiments: JoVE
https://www.readbyqxmd.com/read/27619624/identification-of-b-cell-epitopes-of-borrelia-burgdorferi-outer-surface-protein-c-by-screening-a-phage-displayed-gene-fragment-library
#19
Lucia Pulzova, Zuzana Flachbartova, Elena Bencurova, Lenka Potocnakova, Lubos Comor, Eva Schreterova, Mangesh Bhide
Outer surface protein C (OspC) of Borrelia stimulates remarkable immune responses during early infection and is therefore currently considered a leading diagnostic and vaccine candidate. The sensitivity and specificity of serological tests based on whole protein OspC for diagnosis of Lyme disease are still unsatisfactory. Minimal B-cell epitopes are key in the development of reliable immunodiagnostic tools. Using OspC fragments displayed on phage particles (phage library) and anti-OspC antibodies isolated from sera of naturally infected patients, six OspC epitopes capable of distinguishing between LD patient and healthy control sera were identified...
October 2016: Microbiology and Immunology
https://www.readbyqxmd.com/read/27606593/proteomic-identification-of-immunodiagnostic-antigens-for-trypanosoma-vivax-infections-in-cattle-and-generation-of-a-proof-of-concept-lateral-flow-test-diagnostic-device
#20
Jennifer R Fleming, Lalitha Sastry, Steven J Wall, Lauren Sullivan, Michael A J Ferguson
Trypanosoma vivax is one of the causative agents of Animal African Trypanosomosis in cattle, which is endemic in sub-Saharan Africa and transmitted primarily by the bite of the tsetse fly vector. The parasite can also be mechanically transmitted, and this has allowed its spread to South America. Diagnostics are limited for this parasite and in farm settings diagnosis is mainly symptom-based. We set out to identify, using a proteomic approach, candidate diagnostic antigens to develop into an easy to use pen-side lateral flow test device...
September 2016: PLoS Neglected Tropical Diseases
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