Academic literature on the topic 'Immunodiagnostic assay'
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Journal articles on the topic "Immunodiagnostic assay"
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SAKO, YASUHITO, SONOYO ITOH, MUNEHIRO OKAMOTO, KAZUHIRO NAKAYA, and AKIRA ITO. "Simple and reliable preparation of immunodiagnostic antigens forTaenia soliumcysticercosis." Parasitology 140, no. 13 (June 21, 2013): 1589–94. http://dx.doi.org/10.1017/s0031182013000978.
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SUMMARYCysticercosis caused by infection with the larval stage ofTaenia soliumis an important cause of neurological disease worldwide and immunodiagnosis is important for the control and elimination of cysticercosis. In the present study, we established a simple and reliable preparation of immunodiagnostic low-molecular-weight antigens (LMWAgs) fromT. soliumcyst fluids by a cation-exchange chromatography (CEC). Banding patterns of LMWAgs on SDS-PAGE were different between isolates from Ecuador and China. All cysticercosis patient sera and some echinococcosis patient sera recognized both LMWAgs by enzyme-linked immunosorbent assay (ELISA), but sera from healthy persons were not positive. There was no statistical difference in immunodiagnostic performance of LMWAgs prepared from different geographical isolates. These results indicated that these novel immunodiagnostic antigen preparations could contribute the control and prevention of cysticercosis in endemic areas, especially developing countries.
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Silva, Miriam, Anna Santos, Leticia Rocha, Bruna Caetano, Thais Mitsunari, Luanda Santos, Juliana Polatto, et al. "Development and Validation of Shiga Toxin-Producing Escherichia coli Immunodiagnostic Assay." Microorganisms 7, no. 9 (August 21, 2019): 276. http://dx.doi.org/10.3390/microorganisms7090276.
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Shiga toxin (Stx)–producing Escherichia coli (STEC) and its subgroup enterohemorrhagic E. coli are important pathogens involved in diarrhea, which may be complicated by hemorrhagic colitis and hemolytic uremic syndrome, the leading cause of acute renal failure in children. Early diagnosis is essential for clinical management, as an antibiotic treatment in STEC infections is not recommended. Previously obtained antibodies against Stx1 and Stx2 toxins were employed to evaluate the sensitivity and specificity of the latex Agglutination test (LAT), lateral flow assay (LFA), and capture ELISA (cEIA) for STEC detection. The LAT (mAb Stx1 plus mAb stx2) showed 99% sensitivity and 97% specificity. Individually, Stx1 antibodies showed 95.5% and 94% sensitivity and a specificity of 97% and 99% in the cEIA and LFA assay, respectively. Stx2 antibodies showed a sensitivity of 92% in both assays and a specificity of 100% and 98% in the cEIA and LFA assay, respectively. These results allow us to conclude that we have robust tools for the diagnosis of STEC infections.
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Ogbonna, G., E. Hryhorenko, J. Parsells, S. Phonethepswath, Y. Huang, B. Smith, and L. Sprague. "Performance evaluation of the VITROS® immunodiagnostic products B·R·A·H·M·S PCT assay on the VITROS 3600 immunodiagnostic system." Clinica Chimica Acta 493 (June 2019): S549—S550. http://dx.doi.org/10.1016/j.cca.2019.03.1154.
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Caceres, Diego H., and Mark D. Lindsley. "Comparison of Immunodiagnostic Assays for the Rapid Diagnosis of Coccidioidomycosis in Dogs." Journal of Fungi 8, no. 7 (July 13, 2022): 728. http://dx.doi.org/10.3390/jof8070728.
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Coccidioidomycosis is a disease caused by the dimorphic fungi Coccidioides spp., which affects humans and a variety of animal species, including domestic dogs. In dogs, accurate diagnosis could provide a substantial improvement on the quality of canine life, as well as an advancement in the mapping of regions endemic for coccidioidomycosis. The purpose of this study was to compare immunodiagnostic assays for anti-Coccidioides antibody (Ab) detection in dogs’ serum. Three commercially available immunodiagnostic assays (IMMY®; Norman, OK, USA) were evaluated, including the sōna Coccidioides Ab Lateral Flow Assay (LFA), Coccidioides IDCF immunodiffusion assay (IDCF), and the Clarus Coccidioides Ab Enzyme Immunoassay (EIA). Assays were evaluated using 98 dog serum samples: 29 from dogs with coccidioidomycosis, 15 from dogs diagnosed with histoplasmosis, 10 from dogs diagnosed with blastomycosis, and 44 from dogs without a fungal disease. Using specimens from dogs with coccidioidomycosis, the IDCF had an accuracy of 92% (95% confidence interval [95% CI] = 85–96%), the EIA had an accuracy of 91% (95% CI = 83–96%), and the LFA displayed an accuracy of 82% (95% CI = 73–89%). Using Kappa analysis, the agreement between LFA and EIA was 0.59 (95% CI = 0.42–0.75), that between LFA and IDCF was 0.64 (95% CI = 0.48–0.79), and that between EIA and IDCF was 0.79 (95% CI = 0.64–0.90). Most cross-reactions were observed in dogs with histoplasmosis. Compared with EIA and IDCF, the LFA requires substantially less laboratory equipment and infrastructure and rapidly produces results, offering a substantial improvement for the initial screening of coccidioidomycosis in dogs.
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Procopiou, Michel, Hazel Finney, Scott A. Akker, Shern L. Chew, William M. Drake, Jacky Burrin, and Ashley B. Grossman. "Evaluation of an enzyme immunoassay for plasma-free metanephrines in the diagnosis of catecholamine-secreting tumors." European Journal of Endocrinology 161, no. 3 (September 2009): 509. http://dx.doi.org/10.1530/eje-09-0172e.
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The authors and journal apologize for the error in the above paper which appeared in 161 (1) 131–140. The kits for the assay of free metanephrines were supplied by both Immunodiagnostic Systems Ltd (IDS), Tyne and Wear, UK and also Biotech-IgG (UK) Ltd, Wilmslow, UK.
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Sako, Yasuhito, Minoru Nakao, Takashi Ikejima, Xian Zhi Piao, Kazuhiro Nakaya, and Akira Ito. "Molecular Characterization and Diagnostic Value ofTaenia solium Low-Molecular-Weight Antigen Genes." Journal of Clinical Microbiology 38, no. 12 (2000): 4439–44. http://dx.doi.org/10.1128/jcm.38.12.4439-4444.2000.
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Neurocysticercosis (NCC) caused by infection with the larvae ofTaenia solium is an important cause of neurological disease worldwide. In order to establish an enzyme-linked immunosorbent assay (ELISA) for this infection using recombinant proteins, we carried out molecular cloning and identified four candidates as diagnostic antigens (designated Ag1, Ag1V1, Ag2, and Ag2V1). Except for Ag2V1, these clones could encode a 7-kDa polypeptide, and Ag2V1 could encode a 10-kDa polypeptide. All of the clones were very similar. Except for Ag2V1, recombinant proteins were successfully expressed using anEscherichia coli expression system. Immunoblot analysis of NCC patient sera detected recombinant proteins, but because reactivity to recombinant Ag1 was too weak, Ag1 was not suitable as an immunodiagnostic antigen. So, Ag1V1 and Ag2 were chosen as ELISA antigens, and the Ag1V1/Ag2 chimeric protein was expressed. Of 49 serum samples from NCC patients confirmed to be seropositive by immunoblot analysis, 44 (89.7%) were positive by ELISA. No assays of serum samples from patients with other parasitic infections recognized the Ag1V1/Ag2 chimeric protein. The Ag1V1/Ag2 chimeric protein obtained in this study had a high value for differential immunodiagnosis.
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Ogbonna, G., S. Alvey, E. Hryhorenko, J. Parsells, S. Phonethepswath, and B. Smith. "M142 Analytical and clinical performance of the vitros® immunodiagnostic products B·R·A·H·M·S PCT assay on the vitros immunodiagnostic systems." Clinica Chimica Acta 530 (May 2022): S268. http://dx.doi.org/10.1016/j.cca.2022.04.024.
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Candolfi, E., R. Ramirez, M. P. Hadju, C. Shubert, and J. S. Remington. "The Vitek immunodiagnostic assay for detection of immunoglobulin M toxoplasma antibodies." Clinical and diagnostic laboratory immunology 1, no. 4 (1994): 401–5. http://dx.doi.org/10.1128/cdli.1.4.401-405.1994.
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Prasad, B. V. Siva, Vishal Khatri, P. Suresh Yadav, M. Subhosh Chandra, D. Vijaya Lakshmi, and Kalyan Goswami. "Immunodiagnostic potential ofWuchereria bancroftiL1 antigen–based filarial immunoglobulin G4 detection assay." Transactions of The Royal Society of Tropical Medicine and Hygiene 113, no. 1 (October 15, 2018): 36–43. http://dx.doi.org/10.1093/trstmh/try110.
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Yamasaki, Hiroshi, Takeshi Nakamura, Pewpan M. Intapan, Wanchai Maleewong, Yasuyuki Morishima, Hiromu Sugiyama, Hiroyuki Matsuoka, Kaoru Kobayashi, Katsuyoshi Takayama, and Yukuharu Kobayashi. "Development of a Rapid Diagnostic Kit That Uses an Immunochromatographic Device To Detect Antibodies in Human Sparganosis." Clinical and Vaccine Immunology 21, no. 9 (July 2, 2014): 1360–63. http://dx.doi.org/10.1128/cvi.00149-14.
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ABSTRACTA diagnostic kit using an immunochromatographic device was developed to replace the time-consuming immunodiagnostic methods for human sparganosis. The kit was found to be faster and easier to use than an enzyme-linked immunosorbent assay (ELISA) and showed higher sensitivity and specificity. It will be useful for the laboratory diagnosis of hospitalized cases of sparganosis.
Dissertations / Theses on the topic "Immunodiagnostic assay"
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MAZZOLENI, ELISA. "Exploration of new techniques for purification and chemo-selective conjugation of bioreagents for immunodiagnostic applications." Doctoral thesis, Università degli Studi di Milano-Bicocca, 2015. http://hdl.handle.net/10281/68468.
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Antigene e anticorpo sono i due reagenti chiave di un saggio immunodiagnostico. L’indagine di nuove tecniche e il miglioramento di processi quali la purificazione e la marcatura sito-specifica di antigeni e anticorpi possono promuovere lo sviluppo di nuovi reagenti più efficienti capaci di migliorare la performance dei saggi immunodiagnostici.
La prima parte di questa tesi è stata focalizzata sull’esplorazione di tecniche biotecnologiche innovative nella produzione di antigeni al fine di migliorare i saggi per la rilevazione degli anticorpi IgM e IgG specifici per il virus Epstein-Barr. Il virus EBV è causa della mononucleosi infettiva ed è associato ad un crescente numero di tumori; per questa motivo è importante sviluppare saggi diagnostici per la rilevazione di EBV ad alta specificità e sensibilità. La proteina VCA p18 è uno degli antigeni più importanti per la diagnosi di EBV. I saggi attuali Diasorin LIAISON EBV VCA IgM and IgG si basano su un singolo antigene corrispondente alla regione C-terminale immunodominante della proteina p18, immobilizzata su fase solida. I vari metodi esplorati in questa tesi hanno permesso di ottenere diverse varianti dell’antigene p18 con lo scopo di migliorare le prestazioni dei saggi EBV VCA IgM e IgG a diversi livelli: 1_produzione dell’antigene p18; 2_immobilizzazione dell’antigene p18 su fase solida; 3_formato di saggio. 1_La lunghezza della regione C-terminale della proteina p18 (57aa), risulta essere considerevole per il processo sintetico ma, allo stesso tempo, troppo piccola per essere prodotta in modo efficiente per via ricombinante. Per superare questo problema, abbiamo esplorato il sistema Elastin Like Polypeptides (ELP)-Inteina basato sull’uso di una proteina capace di effettuare auto-cleavage (inteina) e un tag responsivo alla temperatura (ELP). Questa tecnica si è rivelata un eccellente sistema per la produzione del peptide p18. 2_Lo sviluppo di diverse varianti dell’antigene p18 ha permesso di esplorare varie tecniche per l’immobilizzazione dello stesso antigene su fase solida: coating covalente diretto, attraverso il complesso streptavidina-biotina e attraverso l’uso dei peptidi leucine zipper (o velcro). L’immobilizzazione del peptide p18 su fase solida attraverso questi vari metodi è avvenuta con successo e l’attività immunochimica dell’antigene, immobilizzato con queste tecniche innovative, è comparabile o migliore rispetto a quella del peptide sintetico utilizzato nei saggi attuali. 3_Nonostante il saggio Diasorin LIAISON EBV VCA IgM abbia una buona performance analitica, al fine di ottenere un aumento di specificità, è stato esplorato un nuovo tipo di formato di saggio. Sfortunatamente i risultati indicano che questo diverso tipo di formato raggiunge un livello di specificità minore rispetto a quello del saggio attuale.
La seconda parte di questa tesi è stata focalizzata sull’esplorazione di un metodo innovativo per la marcatura sito specifica degli anticorpi. Uno degli approcci più promettenti è basato sulla generazione di gruppi tiolo liberi attraverso la riduzione parziale e selettiva dei ponti disolfuro inter-catena presenti a livello della “hinge region” e la loro reazione con molecole marcanti caratterizzate dal possedere gruppi funzionali reattivi verso i gruppi sulfidrilici. Questa tecnologia è stata utilizzata per la biotinilazione di due diversi anticorpi usati attualmente per il rilevamento della proteina virale p24 di HIV e per l’antigene FGF23. I risultati suggeriscono che la biotinilazione sito-specifica rispetto a quella classica random promuove un miglioramento dell'attività immunochimica degli anticorpi con una conseguente ottimizzazione della performance dei saggi immunodiagnostici.Antigen and antibody are the two key reagents for an immunodiagnostic assay. Investigation of new techniques and improvement of processes such as purification and site-specific labeling of antigen and antibody molecules can promote the development of new more powerful bioreagents able of improving the performance of immunodiagnostic assays. The first part of this thesis aimed to explore innovative biotechnology techniques in antigen production for the improvement of immunoassays that allow the detection of antibodies directed against the Epstein-Barr virus. EBV virus is the causative agent of infectious mononucleosis and it is considered to be associated with a still increasing number of tumors; for this reason it is important to develop diagnostic assays for EBV detection with high specificity and sensitivity. The viral capsid protein VCA p18 is one of the most important antigens for the diagnosis of EBV. The current Diasorin LIAISON EBV VCA IgM and IgG assays rely on a single antigen, consisting in a synthetic peptide corresponding to the immunodominant C-terminal portion of the p18 protein, which is immobilized on solid phase (indirect format). The several methods explored in this thesis have allowed to obtain different variants of the p18 antigen with the aim to improve the performance of DiaSorin LIAISON EBV VCA IgM and IgG assays at different levels: 1_production of p18 antigen; 2_immobilization of p18 antigen on solid phase; 3_immunoassay format. 1_ The length of the immunodominant C-terminal portion of the p18 protein (57aa) appears to be considerable for the synthetic route but, at the same time, too small to be effectively produced in a recombinant fashion. To overcome this problem, we explored the Elastin Like Polypeptides (ELP)-Intein system, a method based on the use of a self-cleavable protein (the intein) and a temperature responsive tag (ELP). This technique has proved to be an excellent system for the preparation of the p18 peptide. 2_The development of different variants of p18 antigen has enabled to explore different techniques for the immobilization of the same antigen on solid phase: direct covalent coating, through streptavidin-biotin complex and through an innovative procedure based on the use of leucine zipper (or “velcro”) peptides. The immobilization of the p18 peptide on solid phase through these different methods occurred successfully and the immunochemical activity of the antigen, immobilized with these innovative techniques, is comparable or better than that of the synthetic peptide used in the current immunoassays. 3_Despite the DiaSorin LIAISON EBV VCA IgM immonoassay has a good analytical performance, in order to obtain an increase of specificity, a new assay format was explored. Unfortunately, the results indicate that this different type of format reaches a lower level of specificity than that of current assay. The second part of this thesis aimed to explore an innovative method for the site-specific labeling of antibodies. One the most promising approach is based on the generation of free thiol groups by selective partial reduction of the interchain disulfide bridges present at the level of the “hinge region” and their reaction to labels carrying sulfhydryl-reactive chemical groups. This technology was used for the biotinylation of two different antibodies currently used in immunoassays for the HIV p24 viral protein and for FGF23 antigen detection. The results suggest that the site-specific biotinylation compared to the random traditional biotinylation promotes a great improvement of antibodies immunochemical activity with a consequent optimization of immunodiagnostic assays performance.
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MENTO, ALFREDO. "Unconventional purification and labelling strategies of bioreagents for immunodiagnostic assays." Doctoral thesis, Università degli Studi di Milano-Bicocca, 2021. http://hdl.handle.net/10281/309986.
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Gli antigeni e gli anticorpi sono reagenti chiave per lo sviluppo di test immunodiagnostici accurati, riproducibili e sensibili, ampiamente utilizzati per la rilevazione di malattie infettive (HIV, HBV, HCV, ecc.) e per la determinazione di marcatori biologici (vitamine, ormoni, ecc.). Questi bioreagenti devono essere prodotti con un alto grado di purezza, in formulazioni stabili e con sufficiente riproducibilità nel tempo (consistenza da lotto a lotto). Un aspetto spesso trascurato nella produzione di questi reagenti è il loro costo, che deve essere sufficientemente basso da non avere un impatto significativo sul prezzo finale dei saggi immunochimici. Le fasi di purificazione e di marcatura di questi bioreagenti incidono principalmente sul loro costo complessivo poiché vengono utilizzati reagenti e strumentazioni costosi e protocolli complessi che richiedono tempo. Per tutti questi motivi è importante ricercare nuove strategie di purificazione che permettano lo sviluppo di processi più semplici, con minor quantità di reagenti, in tempi più brevi, in modo da ridurre i costi. Pertanto, è necessario sviluppare purificazioni innovative e protocolli di marcatura sito-specifici. Nella prima parte di questo progetto abbiamo sfruttato il sistema ELP-intein. Questo metodo si basa sulla combinazione di due tools tecnologici, Elastin-like-polypeptides (ELP) (tool fisico-chimico) e l'attività dell’inteina MxeGyrA (tool biochimico), appartenente alla famiglia delle cis inteine. Ci siamo concentrati sulla purificazione e la marcatura dell'antigene C33 appartenente al virus dell'epatite C (HCV). L'antigene C33, attualmente utilizzato nel test Diasorin LIAISON® XL Murex HCV per la rilevazione di anticorpi umani contro il virus dell'epatite C, è stato purificato utilizzando un metodo non convenzionale senza passaggi cromatografici. Inoltre, abbiamo realizzato una biotinilazione sito-specifica dell'antigene C33 al suo C-terminale durante la purificazione, sfruttando l'attività biologica inteina di MxeGyrA. Sono stati sviluppati due diversi protocolli; entrambi hanno portato all'ottenimento di un antigene C33 biotinilato con un’elevata purezza e immunoreattività paragonabile a quella dell’antigene attualmente utilizzato nel saggio Diasorin HCV. Alla luce di questi buoni risultati, nella seconda parte del progetto, abbiamo studiato la possibilità di applicare la tecnologia del Protein Trans Splicing (PTS) per eseguire la marcatura sito-specifica di bioreagenti. La tecnologia PTS sfrutta l'attività delle split inteine. In particolare, nei nostri esperimenti abbiamo utilizzato la Cfa split-intein che deriva da un processo di mutagenesi della split-intein naturale Npu che ne ha notevolmente migliorato la cinetica di PTS, stabilità termica e tolleranza agli agenti caotropici. Questa nuova tecnica ci ha permesso di impostare un protocollo di marcatura sito-specifica per la produzione di bioreagenti biotinilati. Sono state utilizzate due proteine modello: lo stesso antigene C33 e una IgG umana ricombinante. Anche l'uso della tecnica PTS ha permesso di ottenere per entrambe le due proteine un'elevata purezza e prestazioni comparabili nei relativi saggi immunodiagnostici. In sintesi, il sistema ELP-inteina ha permesso di purificare l'antigene C33 senza passaggi cromatografici e di marcare in modo sito-specifico la stessa proteina al C-terminale. Inoltre, attraverso l'utilizzo del sistema Cfa split-intein abbiamo ottenuto la biotinilazione sito-specifica dell'antigene C33 e dell'IgG ricombinante. Un aspetto molto rilevante è che tutte queste proteine sono funzionali nella piattaforma LIAISON. In futuro, questi protocolli potrebbero essere utilizzati per la purificazione e / o la marcatura sito-specifica di nuovi bioreagenti utili per lo sviluppo di saggi immunodiagnostici.Antigens and antibodies are key reagents for the development of accurate, reproducible and sensible immunodiagnostic assays, which are widely used for the detection of infectious diseases (HIV,HBV, HCV, etc.) and the determination of biological markers (vitamins, hormones, etc.). These bioreagents need to be produced at a high purity degree, in stable formulations and with sufficient reproducibility over time (lot to lot consistency). An aspect often overlooked in the production of these reagents is their cost, which must be low enough to not have a significant impact on the final price of the immunochemical assays. The purification and labeling steps of these bioreagents mainly affect the overall cost of them since costly reagents and instrumentations and complex and time-consuming protocols are used.For all these reasons it is important to seek new purification strategies that allow the development of simpler processes, with less use of reagents and shorter protocol times, and at the end minor costs. Therefore, it is necessary to develop innovative purifications and site-specific labelling protocols. In the first part of this project we exploited the ELP-intein system. This method is based on the combination of two technological tools, the Elastin-like-polypeptides (ELPs) (a physico-chemical tool) and the MxeGyrA intein activity (a biochemical tool), belonging to the cis intein family. We focused on the purification and labelling of the C33 antigen from Hepatitis C Virus (HCV). The C33 antigen, which is currently used in the Diasorin LIAISON® XL Murex HCV assay for the detection of human antibodies against the Hepatitis C Virus, was purified using a not conventional purification method without chromatographic steps. Moreover, we realized a site-specific biotinylation of C33 antigen at its C-terminus during the purification exploiting the MxeGyrA intein biological activity. Two different protocols were developed; both of them brought to the obtainment of a biotinylated C33 antigen with high purity and a comparable immunoreactivity with the one currently used in the Diasorin LIAISON® XL Murex HCV assay. In light of these good results, in the second part of the project, we investigated the possibility to apply the Protein Trans Splicing (PTS) technology to perform site-specific labelling of bioreagents. PTS technology exploits the split intein activity. In particular, in our experiments we used the Cfa split-intein which derives from a mutagenesis process of the natural Npu split-intein that significantly improved its kinetic of PTS, thermal stability and tolerance at the chaotropic agents. This new technique allowed us to set up a site-specific labelling protocol for the production of biotinylated bioreagents. Two model protein were used: the same C33 antigen and a recombinant human IgG. Also the use of PTS technique permitted to obtain for both of two proteins a high purity and a comparable performance in the immunoassays. In summary, the ELP-intein system allowed to purify the C33 antigen without chromatographic steps and then to site-specific label the same protein at the C-terminus. Moreover, through the use of the Cfa split-intein system we obtained the site-specific biotinylation of the C33 antigen and the recombinant IgG. A very relevant aspects is that all these proteins are functional in the LIAISON platform. In the future, these protocols could be used for the purification and-or the site-specific labelling of new bioreagents useful for the development of immunodiagnostic assays.
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Xu, Mengyun. "Optimised label-free biomarker assays with electrochemical impedance spectroscopy." Thesis, University of Oxford, 2013. http://ora.ox.ac.uk/objects/uuid:e527a06b-25e5-48fe-8be5-3c0c10210b74.
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There is huge academic interest and clinical need associated with the development of biomarker immunoassays where general aims are the generation of highly specific, convenient and sensitive sensing formats. In this project, a powerful electrochemical technique, electrochemical impedance spectroscopy (EIS), is applied in the establishment of powerful biomarker detecting protocols. Firstly, ultrasensitive, label-free and reusable insulin sensors, based on an antibody-PEGylated thiol self-assembly monolayer (PEG thiol SAM) interface, were produced and characterised via Faradaic EIS, presenting a detection limit (LOD) of 1.2 pM, a linear range across four orders of magnitude, and high sensitivity in even 50 % serum. By applying similar surface chemistry, a label-free biosensor, specific for the detection of α-synuclein antibodies, was fabricated. The α-synuclein interfaces used enabled the reliable detecting of this biomarker in patient sample serum. The concentration levels in the control and a patient group were determined to be significantly different, and, significantly, this difference was consistently across two different cohorts. Strikingly, this could potentially underpin an entirely new means of early Parkinson’s disease (PD) diagnosis. Non-Faradaic EIS methods were additionally applied to label-free insulin assays at both PEG thiol SAM and zwitterionic polymer film interfaces. The latter presented not only an exceptionally non-fouling interface, but also one seemingly both highly biocompatible and facilitating enhanced receptor: target binding. Finally, impedance assays, though potent, generally, operate by sampling only one of a limited number of available experimental variables, typically, Rct for Faradaic EIS, or C or Z for non-Faradaic EIS. Work carried out herein also explores the generation and utility of a portfolio of mathematically derived immittance functions all obtained from the same raw data sets. A particular focus was the examination of whether these were capable of increasing assay sensitivity and efficiency above normal impedance treatments.
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Leidl, Leonhard [Verfasser]. "Relationship of immunodiagnostic assays for tuberculosis and numbers of circulating CD4+ T-cells in HIV-infection / Leonhard Leidl." Lübeck : Zentrale Hochschulbibliothek Lübeck, 2011. http://d-nb.info/1015754740/34.
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Corral, Marcelo Andreetta. "Imunodiagnóstico da estrongiloidíase humana frente a diferentes frações antigênicas de Strongyloides venezuelensis." Universidade de São Paulo, 2014. http://www.teses.usp.br/teses/disponiveis/5/5134/tde-27082014-111103/.
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A estrongiloidíase é a infecção parasitária causada pelo nematódeo Strongyloides stercoralis. O diagnóstico definitivo é realizado pela visualização de larvas, principalmente nas fezes. Porém as técnicas parasitológicas têm baixa sensibilidade. As técnicas sorológicas apresentam-se como importante alternativa diagnóstica. Pesquisas apontam para a utilização de antígenos heterólogos solúveis, principalmente de Strongyloides venezuelensis. A identificação e caracterização dos antígenos de membrana podem fornecer fonte alternativa de antígenos e assim auxiliar o desenvolvimento das técnicas imunológicas. O presente trabalho teve como objetivo a avaliação das técnicas ELISA e WB frente a diferentes frações antigênicas de larvas filarioides de S. venezuelensis. Foram utilizadas amostras de sangue e fezes de 92 indivíduos, 20 indivíduos com estrongiloidíase (grupo I), 32 indivíduos com outras parasitoses (grupo II) e 40 indivíduos negativos (grupo III) pelos métodos de Lutz, cultura em placa de ágar e Rugai. Para preparação dos antígenos foram utilizadas larvas infectantes obtidas a partir de ratos infectados experimentalmente com S. venezuelensis. Seis frações antigênicas foram preparadas: frações salinas solúveis e de membrana (PBS 0,01M pH 7,2 e SDS 1%, SS e MS; Tris-HCl 25mM pH 7,5 e CHAPS 1%, ST e MT, respectivamente) e frações alcalinas solúvel e de membrana (NaOH 0,15M e SDS 1%, SA e MA, respectivamente). Para a técnica ELISA foram utilizadas placas sensibilizadas com 10ug/mL de antígeno, soro dos indivíduos diluídos 1:200 em PBS 0,05% Tween 3% de leite (PBSTM) e o conjugado (anti IgG-humana peroxidase) em PBSTM. As amostras foram consideradas positivas quando o Índice ELISA foi maior que 1. Para a técnica de WB os soros foram diluídos 1:100 em Tris-HCl 5% de leite (TM) e o conjugado (anti IgG-humana peroxidase) em TM. Após as técnicas sorológicas foram determinadas os parâmetros de diagnóstico pela curva ROC como sensibilidade (SE), especificidade (ES), Likelihood ratio (LR) além da determinação da acurácia diagnóstica (AC) e do índice Kappa (k). A técnica ELISA destacou as frações de membrana com melhor desempenho em relação aos parêmetros diagnósticos estudados (SE 95%, ES 94,4%, AC 94,8%, LR 17,1, k 0,848). O WB revelou componentes antigênicos imunodominantes variando de 260-10kDa, mas destacam-se as frações de 40-35kDa mais frequentes em todas frações antigênicas. Pela técnica de WB, a fração ST apresentou melhor desempenho em relação aos parêmetros diagnósticos estudados (SE 100%, ES 93,1%, AC 94,5, LR 14,4,k 0,854). A utilização das frações de membrana no imunodiagnóstico da estrongiloidíase humana torna-se fonte acessível e eficaz em relação às frações purificadas, não necessitando de gastos complementares para sua obtençãoStrongyloidiasis is a parasitic infection caused by a nematode Strongyloides stercoralis. The definitive diagnosis is made by the larvae visualization in stool samples. However parasitological techniques have low sensitivity. Serological techniques became as suitable diagnostic alternative. Research indicates for the soluble heterologous antigen utilization, mainly Strongyloides venezuelensis. Identification and characterization of membrane antigen may constitute an alternative source of antigen and then assist the development of serological techniques. The aim of this study was evaluate ELISA and WB techniques behind different antigenic fractions of S. venezuelensis´ infective larvae. A total of 92 serum and stool samples was analyzed, 20 from individuals with strongyloidisis (group 1), 32 with other parasitic diseases (group 2) and 40 from individuals with negative coproparasitology (group 3) using Lutz, agar plate culture and Rugai methods. For the antigen preparation infective larvae of S. venezuelensis from experimental infected rats were employed. Six antigenic fractions were prepareted: saline soluble and from membrane fractions (0.01M PBS pH 7.2, and 1% SDS, SS and MS; 25mM Tris-HCl pH 7.5, 1% CHAPS, MT and ST, respectively) and alkaline soluble and membrane fractions (0.15 M NaOH and 1% SDS, SA and MA, respectively). For ELISA technique, plates were sensitized with 10 ug/mL of antigen, serum samples were diluted 1:200 in 0.05% Tween in PBS 3% milk (PBSTM) and conjugate (anti-human IgG peroxidase) in PBSTM. Positive samples were considered when ELISA index was greater than 1. To WB technique, serum samples were diluted 1:100 in Tris-HCl 5% milk (TM) and conjugate (anti-human IgG peroxidase) in the TM. After serological techniques diagnostics parameters were determined by ROC curve how sensitivity (SE), specificity (ES), Likelihood ratio (LR) and determination of diagnostic accuracy (AC) and Kappa (k) index. ELISA technique highlighted the membrane fractions with better performance compared to parameters diagnoses studied (95% SE, 94.4% ES, 94.8% AC, 17.1 LR, 0.848 k). The WB revealed immunodominant antigenic components ranging from 260-10kDa, but there are the fractions of 40-35kDa more frequent in all antigenic fractions. WB technique showed ST fraction better performance in relation to the diagnostic parameters (100% SE, 93.1% ES, 94.5% AC, 14.4 LR, 0.854 k). Membrane fractions in the immunodiagnosis of human strongyloidiasis become an accessible and effective source of antigens in relation to the purified fractions, requiring no additional expense to obtain it
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Gottardi, Maiara. "Imunodiagnóstico da infecção por Strongyloides stercoralis em pacientes candidatos a transplante." Universidade de São Paulo, 2014. http://www.teses.usp.br/teses/disponiveis/5/5134/tde-09012015-162127/.
Full textAbstract:
A estrongiloidíase é uma infecção intestinal causada pelo nematódeo Strongyloides stercoralis. A maioria dos casos evolui para um quadro crônico benigno; entretanto pode haver hiperinfecção e disseminação, sobretudo em pacientes imunodeprimidos. Os métodos parasitológicos convencionais apresentam baixa sensibilidade e, com isso, os testes sorológicos podem representar uma boa alternativa para o diagnóstico dessa helmintíase. O presente trabalho tem como objetivo avaliar as técnicas de RIFI, ELISA e WB para o diagnóstico da estrongiloidíase em pacientes candidatos a transplante. Para validação dos testes, foram utilizadas amostras de soros de pacientes imunocompetentes (positivos para S. stercoralis, positivos para outras parasitoses e negativos). Foram utilizadas amostras de fezes e soros de pacientes candidatos a transplante, a saber: 50 para transplante renal; 50 para transplante de fígado; 50 para transplante de medula óssea. As amostras fecais de todos os pacientes foram analisadas pelas técnicas de sedimentação espontânea, Rugai, e cultura em placa de ágar. Para a execução das técnicas sorológicas foram utilizadas como fonte de antígeno larvas filarioides de S. venezuelensis. Para a RIFI, os soros foram diluídos a 1:40 em tampão PBS-TM e o conjugado anti-IgG humano marcado com fluoresceína diluído 1:500 em PBS acrescido de 4% de azul de Evans, sendo realizada a leitura em microscópio de imunofluorescência, utilizando as objetivas de 20X e 40X. Para a técnica ELISA foram utilizadas placas poliestireno sensibilizadas com 10?g de antígeno (frações solúveis salina e alcalina), soro dos indivíduos diluídos a 1:200 em PBS 0,05% Tween 3% de leite (PBS-TM) e conjugado (anti-IgG humana peroxidase) em PBS-TM. As amostras foram consideradas positivas quando o índice ELISA foi maior que 1. Para a técnica de WB, os soros foram diluídos 1:100 em Tris-HCl 5% de leite (TM) e o conjugado (anti IgG-humana peroxidase) em PBS-TM. Dos 150 pacientes candidatos a transplante analisados, 9,3% (n=14) foram positivos pelas técnicas parasitológicas, sendo que a cultura em placa de ágar detectou 6,6% (n=10). Na RIFI a soropositividade para detecção da infecção por S. stercoralis foi de 16,6% (n=25), sendo, 22% transplante renal, 18% hepático e 10% medula óssea. Pela técnica ELISA a soropositividade foi de 11,3% (n=17), sendo 14% nos pacientes candidatos a transplante renal, 14% hepático e 3% medula óssea, utilizando o antígeno alcalino, e 24,6% (n=37), sendo 18% nos pacientes candidatos a transplante renal, 50% hepático e 6% medula óssea, utilizando o antígeno salino. Pela técnica WB a soropositividade foi de 20,6% (n=31), sendo 18% nos pacientes candidatos a transplante renal, 32% hepático e 12% medula óssea, utilizando o antígeno alcalino, e 18,6% (n=28) sendo 10% nos pacientes candidatos a transplante renal, 14% hepático e 32% medula óssea, utilizando o antígeno salino. A utilização de técnicas imunodiagnósticas pode ser indicada na triagem de pacientes em fila de transplante, devendo-se levar em conta as limitações de reações sorológicas em pacientes imunodeprimidosStrongyloidiasis is an intestinal infection caused by the nematode Strongyloides stercoralis. Most cases progress to a benign chronic condition; however hyperinfection and dissemination may occur, especially in immunocompromised patients. Conventional parasitological methods have low sensitivity and, thus, serological tests may be a good alternative for the diagnosis of this helminthiasis. The aim of this study was to evaluate RIFI, ELISA and WB techniques for the diagnosis of strongyloidiasis in patients candidates for transplants. In order to validate the tests samples of sera from immunocompetent patients (positive for S. stercoralis, positive for other parasitosis, and negatives) were used. Feces and sera samples of patients candidates for transplants were used as follows: 50 for renal transplant, 50 for liver transplant, 50 for bone marrow transplant. Fecal samples of all patients were analyzed by spontaneous sedimentation, Rugai and Agar plate culture techniques. Filarioid larvae of S. venezuelensis were used as source of antigen for serological tests. For RIFI, sera were diluted 1:40 in PBS-TM buffer and human anti-IgG conjugate labeled with fluorescein was diluted 1:500 in PBS with 4% Blue Evans, reading being performed in immunofluorescence microscope using 20X and 40X objectives. For ELISA technique polystyrene plates sensitized with 10?g of antigen (saline and alkaline soluble fractions), individual sera diluted at 1:200 in PBS 0.05% Tween 3% of milk (PBS-TM) and conjugate (human anti-IgG peroxidase) in PBS-TM were used. Samples were considered positive when ELISA index was higher than 1. For WB technique, sera were diluted in Tris-HCl 5% of milk (TM) and conjugate (human anti-IgG peroxidase) in PBS-TM. From 150 patients candidates for transplants analyzed 9.3% (n=14) were positive by parasitological techniques, agar plate culture detected 6.6% (n=10). In RIFI seropositivity for S. stercoralis infection detection was 16.6% (n=25) being 22% renal transplant, 18% hepatic and 10% for bone marrow. By ELISA technique seropositivity was 11.3% (n=17), being 14% in patients candidates for renal transplant, 14% hepatic and 3% bone marrow, using alkaline antigen and 24.6% (n=37), being 18% in patients candidates for renal transplant, 50% hepatic and 6% bone marrow, using saline antigen. By WB technique seropositivity was 20.6% (n=31), being 18% in patients candidates for renal transplant, 32% hepatic and 12% bone marrow, using alkaline antigen, and 18.6% (n=28) being 10% in patients candidates for renal transplant, 14% hepatic and 32% bone marrow, using saline antigen. Application of immunodiagnostic techniques could be indicated in screening of patients in transplant waiting list, however limitations of serological reactions in immunodepressed patients should be considered
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Boshoff, Christoffel Hendrik. "Maedi-Visna virus : the development of serum and whole blood immunodiagnostic assays." Thesis, 1997. http://hdl.handle.net/10413/6083.
Full textAbstract:
This thesis describes the development of serum and whole blood immunodiagnostic assays for Maedi-Visna virus (MVV). All previously described recombinant MVV ELISA assays utilised either the core p25 or transmembrane (TM) proteins alone, or combined, but as individual proteins. The p25 and TM genes of MVV were cloned individually into the pGEX-2T expression vector. Both proteins were expressed as a combined fusion protein in frame with glutathione S-transferase (GST). The purified
recombinant antigens (GST-TM and GST-TM-p25) were used to develop a MVV ELISA. Sera from 46 positive and 46 negative sheep were tested using the GST-TM and GST-TM-p25 ELISAs and a commercial p25 EIA kit. A two-graph receiver operating characteristic (TG-ROC) analysis program was used to interpret the data. The GST-TM-p25 ELISA was more sensitive than the commercial assay which is based on the p25 antigen alone and more specific than the GST-TM ELISA. The GST-TM-p25 ELISA showed a sensitivity and specificity of 100%. The human AIDS lentivirus transmembrane (TM) glycoprotein portion of the envelope viral protein has been identified as the antigen most consistently recognised by antibodies. There is suggestive evidence that the same applies to MVV as the GST-TM fusion protein, expressed in E. coli, has comparable sensitivity to the GST-TM-p25 fusion protein, but lacks specificity. However, due to the hydrophobic nature of the MVV TM protein, purification of the expressed fusion protein required lengthy purification protocols. This was despite the fact that only a truncated version of the TM protein was expressed. This prompted investigating an alternative expression system that could possibly circumvent the above mentioned problems. The yeast Pichia pastoris is known to be suitable for the high-level expression of heterologous proteins which are secreted into the culture supernatant. These features made P. pastoris an attractive host for the expression of the hydrophobic TM protein of MVV. However, limited success was achieved as only low expression levels were obtained and detection and quantification was only accomplished by means of ELISA. Evaluation of the diagnostic performance of the P. pastoris expressed MVV TM-polypeptide was performed using a panel of 36 confirmed negative and positive sera, and evaluated using a TG-ROC analysis programme, which yielded an equal Se and Sp of 83%. The use of a novel rapid immunoassay system, which allows the detection of circulating antibodies in whole blood, has been investigated for use as a MVV diagnostic assay. The central feature of this immunoassay lies in a monoclonal antibody against a glycophorin epitope present on all sheep erythrocytes. A Fab'-peptide conjugate was constructed by coupling a synthetic peptide, corresponding to a sequence from MVV TM protein, to the hinge region of the Fab' fragment of the antisheep erythrocyte antibody. Within the limited number of 10 seronegative and 10 seropositive samples the autologous red blood cell agglutination assay had a sensitivity of 90% and a specificity of 80%. Despite the limitations and difficulties encountered, the use of such rapid whole blood immunodiagnostic assays for MVV holds promise.Thesis (Ph.D.)-University of Natal, Durban, 1997.
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YANG, HONG-TONG, and 楊宏通. "Immunodiagnosis of trichomoniasis vaginalis:comparison between indirect fluorescent antibody test (IFAt) and enzyme-linked immunosorbent assay (ELISA)." Thesis, 1989. http://ndltd.ncl.edu.tw/handle/10639050401194443631.
Full textBooks on the topic "Immunodiagnostic assay"
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W, Burgess Graham, and James Cook University of North Queensland. Graduate School of Tropical Veterinary Sciences., eds. ELISA technology in diagnosis and research. Townsville, Australia: Graduate School of Tropical Veterinary Science, James Cook University of North Queensland, 1988.
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Marius, Teodorescu, and Froelich Christopher J, eds. Advanced immunoassays in rheumatology. Boca Raton: CRC Press, 1994.
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N, Khan Masood, and Findlay John W. A, eds. Ligand-binding assays: Development, validation, and implementation in the drug development arena. Hoboken, N.J: Wiley, 2010.
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W, Pruzanski, and Keystone E. C, eds. Paraproteins in disease: Investigation of plasma cell dyscrasia. Edinburgh: Churchill Livingstone, 1985.
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Quantitative Immunoassay: A Practical Guide for Assay Establishment, Troubleshooting, and Clinical Application. AACC Press, 2000.
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Elisa as diagnostic tool: Prospects and implications. New Delhi: Wiley Eastern, 1992.
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Khan, Masood N., and John W. Findlay. Ligand-Binding Assays: Development, Validation, and Implementation in the Drug Development Arena. Wiley & Sons, Incorporated, John, 2009.
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Khan, Masood N., and John W. Findlay. Ligand-Binding Assays: Development, Validation, and Implementation in the Drug Development Arena. Wiley & Sons, Incorporated, John, 2009.
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Findlay, John W. A., and Masood N. Khan. Ligand-Binding Assays: Development, Validation, and Implementation in the Drug Development Arena. Wiley & Sons, Incorporated, John, 2009.
Find full textBook chapters on the topic "Immunodiagnostic assay"
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Pappas, Michael G. "Immunodiagnostic Assays." In The Biotech Business Handbook, 247–72. Totowa, NJ: Humana Press, 1994. http://dx.doi.org/10.1007/978-1-4612-0293-6_17.
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Kinders, R., J. Slota, J. Patrick, C. Plate, I. Kafer, W. Caminiti, H. Rittenhouse, and G. Manderino. "An Assay for Cryptic Tumor Antigens in Sera of Women with Breast Cancer." In Breast Cancer Immunodiagnosis and Immunotherapy, 55–67. Boston, MA: Springer US, 1989. http://dx.doi.org/10.1007/978-1-4757-1296-4_6.
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Ludwig, George V., Cynthia A. Rossi, and Robert L. Bull. "Concepts for the Development of Immunodiagnostic Assays for Detection and Diagnosis of Biothreat Agents." In Biological Weapons Defense, 551–79. Totowa, NJ: Humana Press, 2005. http://dx.doi.org/10.1385/1-59259-764-5:551.
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Takemoto, Y., M. Tsuji, and Y. Iwanaga. "Immunodiagnosis for Anisakiasis with Detection of IgE Antibody to Human Anisakiasis by Enzyme-Linked Immunosorbent Assay." In Intestinal Anisakiasis in Japan, 187–90. Tokyo: Springer Japan, 1990. http://dx.doi.org/10.1007/978-4-431-68299-8_23.
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"Example 3 Troponin measurement by immunoassay – Problem of low assay sensitivity and interference from heterophilic antibodies." In Immunodiagnostics and Patient Safety. Berlin, New York: DE GRUYTER, 2011. http://dx.doi.org/10.1515/9783110249484.129.
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Ricardo, Lujan, and Cupp Eddie W. "Human Onchocerciasis: New Immunodiagnostic Assays and Control Measures." In Biotechnology for Biological Control of Pests and Vectors, 179–92. CRC Press, 2018. http://dx.doi.org/10.1201/9781351070300-17.
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Sadeghalvad, Mona, and Nima Rezaei. "Introduction on Monoclonal Antibodies." In Monoclonal Antibodies. IntechOpen, 2021. http://dx.doi.org/10.5772/intechopen.98378.
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Monoclonal antibodies (mAbs) are a group of antibodies produced by identical clones of B lymphocytes against a particular antigen. mAbs are identical in several properties such as protein sequence, antigen-binding site region, binding affinity for their targets, and identical downstream functional effects. These characteristics of mAbs highlight their differences with the polyclonal antibodies which have heterogenous activities and recognize different epitopes on an antigen. Murine mAbs was the first generation of mAbs developed by hybridoma technology however, because of their murine origin, they can trigger the anti-mouse antibody response in the host which could accelerate mAb clearance and undesirable allergic reactions upon repeated administration. This issue was resolved by developing engineering methods toward producing less immunologic chimeric or humanized antibodies. mAbs applications have become a novel way of targeting antigens in a wide variety of diseases such as autoimmunity, malignancies, and asthma. In addition, high specificity and high affinity binding properties of mAbs make them effective biological reagents in immunodiagnostic assays. They can be used in diagnosis of infectious diseases and detection of certain antigens or in serological assessments for detection of antibodies against a certain antigen. This chapter summarizes the general properties of mAbs, their production processes, and their important diagnostic and therapeutic applications.
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H. Ravindranath, Mepur, and Fatiha E.L. Hilali. "Monospecific and Polyreactive Monoclonal Antibodies against Human Leukocyte Antigen-E: Diagnostic and Therapeutic Relevance." In Monoclonal Antibodies. IntechOpen, 2021. http://dx.doi.org/10.5772/intechopen.95235.
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A monoclonal antibody (mAb) binds to an antigen recognizing an epitope (a sequence of amino acids). A protein antigen may carry amino acid sequence unique to that antigen as well as sequences found in other proteins. Human leukocyte antigens (HLA), a family of proteins expressed by the Major Histocompatibility Complex gene family represent a special case, in that it displays a high degree of polymorphism. Every HLA molecule possesses both specific (private) epitopes and epitopes shared (public) with other HLA class Ia and class Ib molecules. HLA-E is overexpressed in cancer cells more than any other HLA Class I molecules. Therefore specific localization of HLA-E with mAbs is pivotal for developing targeted therapy against cancer. However, the commercially available mAbs for immunodiagnosis are polyreactive. We have developed anti-HLA-E mAbs and distinguished monospecific from polyreactive mAbs using Luminex multiplex single antigen bead (SAB) assay. HLA-E-binding of monospecific-mAbs was also inhibited by E-restricted epitopes. The amino acid sequences in the region of the epitopes bind to CD94/NKG2A receptors on CD8+ T cells and NK cells and block their antitumor functions. Monospecific-HLA-E mAbs recognizing the epitopes sequences can interfere with the binding to restore the anti-tumor efficacy of NK cells. Also, monospecific-mAbs augment the proliferation of CD4-/CD+ cytotoxic T-lymphocytes. Therefore, anti-HLA-E monospecific-mAb can serve as a double-edged sword for eliminating tumor cells.
Conference papers on the topic "Immunodiagnostic assay"
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Essone, Paulin N., Tom H. Ottenhoff, Novel N. Chegou, and Gerhard Walzl. "Tuberculosis Immunodiagnostic Tests: A Potential Shift From Iterferon-Gamma (IFN-³) Release Assays To Tumor Necrosis Factor-Alpha (TNF-±) Release Assays With Newly Identified Mycobacterium Tuberculosis Antigens." In American Thoracic Society 2012 International Conference, May 18-23, 2012 • San Francisco, California. American Thoracic Society, 2012. http://dx.doi.org/10.1164/ajrccm-conference.2012.185.1_meetingabstracts.a6508.
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