DOCSLIB.ORG

Estimation of Igg and Igm Brucella Antibodies in Infected and Non-Infected Persons by a Radioimmune Technique

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Estimation of Igg and Igm Brucella Antibodies in Infected and Non-Infected Persons by a Radioimmune Technique J Clin Pathol: first published as 10.1136/jcp.37.6.692 on 1 June 1984. Downloaded from J Clin Pathol 1984;37:692-696 Estimation of IgG and IgM brucella antibodies in infected and non-infected persons by a radioimmune technique WG HEWITT, DJH PAYNE* From the Public Health Laboratory, St Mary's Hospital, Portsmouth P03 6A Q SUMMARY Sera from 105 blood donors and eight patients with brucellosis were examined for anti-brucella IgG and IgM by a radioimmune technique. A pooled standard was used for com- parison and evaluation. The upper limit of the 99% confidence interval on the mean of both immunoglobulin classes in blood donor sera was below 7 units/ml. Antibody response was shown in three acute, two relapsing, two chronic, and one asymptomatic cases. Values of up to 300 units/ml of both classes were found in the acute cases. Chronic sufferers showed lower concentra- tions of antibody. Relapsing cases showed increases comprising mainly IgG. The assay method, which shows general concordance with the results of conventional tests, is useful and measures individual immunoglobulin classes directly. copyright. Most human cases of brucellosis are diagnosed by cally suffering from the disease. Mercaptoethanol recognising the symptoms and by demonstrating the may be used to differentiate between IgG and IgM antibodies in the serum. The organism may be iso- activity89 but is difficult to control and the results are lated from the blood, but bacteraemia is intermit- not readily reproducible between laboratories. The tent.' Reliance is therefore placed on immunological course of the disease may therefore not be reflected procedures, which must be both accurate and infor- fully in the results of conventional tests. http://jcp.bmj.com/ mative. Titres obtained in complement fixation and serum Conventionally, antibodies have been studied agglutination tests derive from the in vitro activity of using the serum agglutination test, the complement the total antibody population of the serum. Proce- fixation test, and the indirect antihuman globulin dures have been devised recently which have the assisted agglutination test. From these results the advantage of demonstrating the immunoglobulin presence and relative proportions of anti-brucella classes individually. They include enzyme linked immunoglobulins (IgG and IgM) are inferred and immunosorbent assay'0 and immunofluorescence on September 25, 2021 by guest. Protected thus the stage of the disease and its activity in the tests." For some time this laboratory has been patient are estimated.24 examining sera. by means of immunoradiometric Previous workers have shown that the titres assay, and we report here the results obtained with obtained in conventional serological tests are not sera from blood donors and from patients with absolutely related to the presence and relative pro- brucella infection. portions of IgG and IgM. Agglutination may be caused by both IgM and IgG, and complement Material and methods fixation test activity may be high throughout the dis- ease. Acute cases may present with high, low, or SERA undetectable concentrations of antibody.67 High Blood donor sera were obtained from the National titres may be produced in persons, such as vet- Blood Transfusion Service Laboratory at Southamp- erinarians, exposed to the organism but not clini- ton. Standards for comparison were prepared from pools of specimens showing high serum agglutina- *Dr DJH Payne died in April 1982. tion and complement fixation test results. Other sera were those examined by this brucella reference Accepted for publication 5 March 1984 laboratory. All were stored at - 20°C before testing. 692 J Clin Pathol: first published as 10.1136/jcp.37.6.692 on 1 June 1984. Downloaded from Radioimmune estimation of brucella antibodies 693 PREPARATION OF ANTIGEN performed according to the methods of Kerr et al.'5 EEC Standard Brucella abortus antigen (Central Veterinary Laboratory, Weybridge) was centrifuged Results at 2400 g for 30 min and the deposited cells taken up in 0*85% (0.124 mol/1) saline containing 0-4% Histograms of the distributions of the specific IgM (0.043 mol/l) phenol. The packed cell concentration and IgG concentrations of 105 samples of blood was adjusted to 5 % as measured by microhaemato- donor serum are shown in Figs. 1 and 2. If a Poisson crit. distribution can be assumed, the following statistics are obtained. For IgM: mean = 3 11 U/ml, standard PREPARATION OF RADIOACTIVE LABELLED deviation (SD) = 1-42 U/ml. For IgG: mean = 2-47 ANTIHUMAN GLOBULIN and SD = 1-92 U/ml. Ninety nine percent of such Goat antihuman IgM and goat antihuman IgG sera blood donors can therefore be expected to have (Technicon) were labelled with '25Iodine (Amer- concentrations of less than 7 U/ml of both immuno- sham International PLC) by the chloramine T pro- globulin classes. cess.'2 Unbound iodine was removed by gel filtration Forty nine specimens from eight patients sus- using Sephadex G25 coarse grade (Pharmacia Fine pected of suffering from brucellosis were examined Chemicals Ltd). After a series of titrations we found (Table). In many cases the antihuman globulin that 30 ml of labelled antihuman globulin at a assisted agglutination test was also performed. strength suitable for the test could be prepared from the original 2 ml of goat serum. Phosphate buffered saline at pH 7-3 (PBS, Oxoid) was used as diluent 18 and eluent. 16 14 IMMUNORADIOMETRIC ASSAY The method used was similar to that of Parratt et al 3 and Brown and Lee.'4 Fifty microlitres of serum or serum diluted in PBS was added to 200 ul of anti- copyright. gen. After overnight incubation at 37°C unbound proteins were removed by repeated washing in PBS containing 2-5 g/l of bovine albumin (Fraction V, Sigma) using centrifugation at 2000 g at 8°C for 20 min. The spun deposit was then resuspended in 0-2 2 ml of the diluent, 100 ,ul of radiolabelled antihuman globulin (anti-IgG or anti-IgM) was added, and the 0 3 4 5 6 7 8 9 10 11 12 http://jcp.bmj.com/ mixture was incubated at 37°C for 90 min. Unbound IRMA IgM concentration (U/ml) constituents were removed as previously. The Fig. 1 Distribution ofanti-Brucella IgM in 105 blood radioactivity present in the washed deposit was donors. IRMA = immunoradiometric assay. measured using a gamma counter (Wilj model 2001). 18- CALCULATION OF RESULTS 16- on September 25, 2021 by guest. Protected The amount of IgM and IgG as shown by the test was assigned a numerical value by comparison with 14- a serum standard, to which had been assigned an 12 - arbitrary content. This had been adopted after con- c 10- sultation with other workers in the field. Duplicate a.; a 8- tests were performed for both immunoglobulin clas- LA- ses in test sera. A standard graph was prepared from 6- examination of dilutions of the standard serum, each 4- being done in triplicate or quadruplicate. Results and statistical analyses were calculated using a mic- 2- _H_ _n_ rocomputer (CBM). i nm - 0 1 2 3 4 5 6 7 8 9 10 11 12 CONVENTIONAL TESTS IRMA IgG concentration (U/ml) Serum agglutination, complement fixation, and Fig. 2 Distribution ofanti-Brucella IgG in 105 blood antihuman globulin assisted agglutination tests were donors. IRMA = immunoradiometric assay. J Clin Pathol: first published as 10.1136/jcp.37.6.692 on 1 June 1984. Downloaded from 694 Hewitt, Payne Results oftests on sera from eight cases ofbrucellosis Case no Months after onset of SA T CFT IRMA symptoms B ab B mel B ab B mel IgG IgM 1 1 1280 640 >256 128 285 150 4 Is .~~~~~~~~-Is ISx i's 166. 17 5 640 160 256 32 142 13 2 5120 5120 >256 >256 37 321 13 2560 2560 256 128 70 280 1-8 2560 1280 128 32 92 173 3 640 640 32 8 52 50 320 640 256 128 21 132 4 <20 <20 64 64 107 42 2 5120 5120 >256 >256 313 >200 3 1280 1280 >256 >256 223 241 7 640 640 >256 >256 322 73 7-4 640 320 64 >256 241 56 8 640 320 >256 >256 IS IS 10 160 160 AC AC 78 18 -8 <20 <20 <4 <4 IS IS 1 1280 1280 128 128 51 85 1-6 320 320 >256 256 94 46 4 40 <20 8 8 7 4 13 <20 <20 <4 <4 3 2 25 <20 <20 8 4 4 1 28 640 160 >256 >256 104 7 6 (10 years) 80 80 AC AC 28 14 +2 40 40 8 8 18 10 6 40 40 8 8 18 10 11 <20 <20 16 16 IS IS copyright. 17 80 <20 8 16 IS IS 40 <20 <20 16 16 28 13 47 40 40 AC AC 23 10 54 80 40 8 8 19 8 56 80 80 8 8 18 5 60 80 80 16 8 13 4 68 40 80 16 8 11 4 7 (2 years) <20 <20 AC AC IS IS http://jcp.bmj.com/ +1 <20 <20 AC AC 18 10 2 40 <20 4 <4 17 11 4 <20 <20 128 16 59 20 7 160 160 128 64 IS IS 9 160 160 128 64 IS IS 10 80 80 32 32 IS IS 13 80 160 64 32 IS IS 25 80 40 16 8 32 10 8 (No symptoms) <20 <20 <4 <4 IS IS on September 25, 2021 by guest. Protected 14 <20 <20 <4 <4 5 2 27 <20 <20 <4 <4 3 2 36 320 160 256 128 330 32 38 640 320 256 128 159 25 40 160 80 256 64 99 14 49 <20 <20 32 16 27 5 B ab and B mel = B abortus and B melitensis antigen, respectively, used in the test.
Load more

Recommended publications
  • Memoranaurns by the Participants in Signes Par Les Partici- I the Meeting
    Memoranda are state- Les Memorandums ments concerning the exposent les conclu- /e , conclusions or recom- sions et recomman- M e mmooranrantedaa mendations of certain dations de certaines / a t w / /WHO scientific meet- /reunions scientifiques ings; they are signed de /'OMS; ils sont Memoranaurns by the participants in signes par les partici- I the meeting. pants a ces reunions. Bulletin ofthe World Health Organization, 62 (2): 217-227 (1984) © World Health Organization 1984 Immunodiagnosis simplified: Memorandum from a WHO Meeting* Technologies suitable for the development ofsimplified immunodiagnostic tests were reviewed by a Working Group of the WHO Advisory Committee on Medical Research in Geneva in June 1983. They included agglutination tests and use ofartificialparticles coated with immunoglobulins, direct visual detection of antigen-antibody reactions, enzyme- immunoassays, and immunofluorescence and fluoroimmunoassays. The use of mono- clonal antibodies,in immunodiagnosis and of DNA /RNA probes to identify viruses was also discussed in detail. The needfor applicability of these tests at three levels, i.e., field conditions (or primary health care level), local laboratories, and central laboratories, was discussed and their use at thefield level was emphasized. Classical serological techniques have been used for All these tests can be carried out in laboratories that a long time for diagnostic purposes, e.g., for con- are equipped with basic instruments as well as special- firmation of clinical diagnoses, epidemiological ized apparatus (e.g., gamma counters for RIA, ultra- studies, testing of blood donors, etc. Some of these violet microscopes for IMF, etc.), which are usually techniques have been standardized to a high degree of available only in the larger, central laboratories.
    [Show full text]
  • Radioimmunoassay in Developing Countries: General Principles
    XA9847613 Chapter 16 RADIOIMMUNOASSAY IN DEVELOPING COUNTRIES (General principles) R.D. Piyasena Radioimmunoassay (RIA) is probably the most commonly performed nuclear medicine technique. It is an in vitro procedure, where no radioactivity is administered to the patient. But this alone is not the reason for its widespread use. It provides the basis for extremely sensitive and specific diagnostic tests, and its use in present day medicine has brought a virtual information explosion in terms of understanding the pathophysiology of many diseases. The fact that the technology involved is within the technical and economic capabilities of the developing world is evident from the increasing demand for its introduction or expansion of existing services. RIA facilities need not be restricted to urban hospitals, as in the case of in vivo nuclear medicine techniques, but may be extended to smaller district hospitals and other laboratories in peripheral areas. It is also possible to send blood samples to a central laboratory so that a single centre can serve a wide geographical area. There are many laboratories in the industrialized world that receive a major proportion of samples for assay by mail. In recent years, substantial RIA services have been established in many of the developing countries in Asia and Latin America. The International Atomic Energy Agency (IAEA) and World Health Organisations (WHO) have made vital contributions to these activities and have played a catalytic role in assisting member states to achieve realistic goals. In the past five years, more than 250 individual RIA laboratories in developing member states have been beneficiaries of IAEA projects.
    [Show full text]
  • Frequently Asked Questions
    Frequently Asked Questions 03-DFAQ-02a version 1 Contents 1. COMMERCIAL INFORMATION .................................................................................................... 4 1.1. How can I place an order? ...........................................................................................................4 1.2. How can I obtain a price? ............................................................................................................4 1.3. Does DIAsource provide the kit components separately? ...........................................................4 1.4. How can I address a complaint? ..................................................................................................4 2. REGULATORY INFORMATION ..................................................................................................... 4 2.1. Do the DIAsource kits contain dangerous substances? ...............................................................4 2.2. Are the DIAsource assays CE mark and/or FDA approved? .........................................................4 2.3. May the DIAsource assays be sold in Canada? ............................................................................5 3. TECHNICAL INFORMATION ......................................................................................................... 5 3.1. How can I receive a technical information? ................................................................................5 3.2. What is the principle of an immunoassay (ELISA – RIA – IRMA)? ................................................5
    [Show full text]
  • Design, Visualize and Detect
    Thermo Scientific Pierce Antibody Immunostaining Guide Design, visualize and detect Immunostaining Primary Antibodies • Secondary Antibodies • Staining Dyes • Kits Thermo Scientific Table of Contents Pierce Antibody Immunostaining Guide Page Introduction 1-3 The Cell 4-5 Mammalian Cell Type Choices 6-8 Immunohistochemistry 9-12 Immunofluorescence 13-22 Secondary Antibodies 23-27 Primary Antibodies by Cellular Structures 28-31 by Research Areas 32-43 by Cell Signaling 44-59 by Biological Processes 60-76 Left: Detection of mouse anti-α-tubulin in an A549 cell in Telophase with Thermo Scientific DyLight Dye 550-GAM. Chromosomes (orange) at the poles become Introduction diffuse, while nuclei (blue) divide into two future cells. Immunofluorescence (IF) and immunohistochemistry (IHC) are two methods commonly used to detect proteins in a cellular context. Immunofluorescent detection of proteins can be performed on both fixed cells in culture and on paraffin or frozen tissue sections. The advantages of using IF to detect cellular proteins includes the ability to visualize the subcellular location of protein(s) of interest, assess both protein expression and post-translational modifications, and design multiplex experiments. When IF detection is extended to tissues sections (IHC), a higher level of resolution is achieved because researchers are analyzing target protein(s) in a near physiological state, making it ideal for assessing normal and disease tissues. To order, call 800-874-3723 or 815-968-0747. Outside the United States, contact your local branch office or distributor. 1 Need Antibodies? Build a Better Antibody Introduction We have over 30,000 antibodies in 42 research areas. Use our custom services to produce antibodies you can trust.
    [Show full text]
  • A New Human Chromogranin a (Cga) Immunoradiometric Assay Involving Monoclonal Antibodies Raised Against the Unprocessed Central Domain (145–245)
    British Journal of Cancer (1999) 79(1), 65–71 © 1999 Cancer Research Campaign A new human chromogranin A (CgA) immunoradiometric assay involving monoclonal antibodies raised against the unprocessed central domain (145–245) F Degorce1, Y Goumon2, L Jacquemart1, C Vidaud1, L Bellanger1, D Pons-Anicet1, P Seguin1, MH Metz-Boutigue2 and D Aunis2 1CIS biointernational, Division In Vitro Technologies, BP175, 30203 Bagnols-sur-Cèze, France; 2Institut National de la Santé et de la Recherche Médicale, INSERM U-338, Biologie de la Communication Cellulaire, 67084 Strasbourg, France Summary Chromogranin A (CgA), a major protein of chromaffin granules, has been described as a potential marker for neuroendocrine tumours. Because of an extensive proteolysis which leads to a large heterogeneity of circulating fragments, its presence in blood has been assessed in most cases either by competitive immunoassays or with polyclonal antibodies. In the present study, 24 monoclonal antibodies were raised against native or recombinant human CgA. Their mapping with proteolytic peptides showed that they defined eight distinct epitopic groups which spanned two-thirds of the C-terminal part of human CgA. All monoclonal antibodies were tested by pair and compared with a reference radioimmunoassay (RIA) involving CGS06, one of the monoclonal antibodies against the 198–245 sequence. It appears that CgA C-terminal end seems to be highly affected by proteolysis and the association of C-terminal and median-part monoclonal antibodies is inadequate for total CgA assessment. Our new immunoradiometric assay involves two monoclonal antibodies, whose contiguous epitopes lie within the median 145–245 sequence. This assay allows a sensitive detection of total human CgA and correlates well with RIA because dibasic cleavage sites present in the central domain do not seem to be affected by degradation.
    [Show full text]
  • Diagnosis of Brucellosis by Serology Klaus Nielsen* Animal Diseases Research Institute, 3851 Fallow®Eld Road, Nepean, Ont., Canada K2H 8P9
    Veterinary Microbiology 90 (2002) 447±459 Diagnosis of brucellosis by serology Klaus Nielsen* Animal Diseases Research Institute, 3851 Fallow®eld Road, Nepean, Ont., Canada K2H 8P9 Abstract Serological diagnosis of brucellosis began more than 100 years ago with a simple agglutination test. It was realized that this type of test was susceptible to false positive reactions resulting from,for instance,exposure to cross reacting microorganisms. It was also realized that this test format was inexpensive,simple and could be rapid,although results were subjectively scored. Therefore,a number of modi®cations were developed along with other types of tests. This served two purposes: one was to establish a rapid screening test with high sensitivity and perhaps less speci®city and a con®rmatory test,usually more complicated but also more speci®c,to be used on sera that reacted positively in screening tests. This led to another problem: if a panel of tests were performed and they did not all agree,which interpretation was correct? This problem was further compounded by the extensive use of a vaccine which gave rise to an antibody response similar to that resulting from ®eld infection. This led to the development of an assay that could distinguish vaccinal antibody,starting with precipitin tests. These tests did not perform well,giving rise to the development of primary binding assays. These assays,including the competitive enzyme immunoassay and the ¯uorescence polarization assay are at the apex of current development,providing high sensitivity and speci®city as well as speed and mobility in the case of the ¯uorescence polarization assay.
    [Show full text]
  • Detailed Follow-Up of Hepatitis B Surface Antigen Positive Blood Donors
    Open Research Online The Open University’s repository of research publications and other research outputs Detailed follow-up of hepatitis B surface antigen positive blood donors. Thesis How to cite: Brown, Susan (1992). Detailed follow-up of hepatitis B surface antigen positive blood donors. The Open University. For guidance on citations see FAQs. c 1992 The Author https://creativecommons.org/licenses/by-nc-nd/4.0/ Version: Version of Record Link(s) to article on publisher’s website: http://dx.doi.org/doi:10.21954/ou.ro.00010184 Copyright and Moral Rights for the articles on this site are retained by the individual authors and/or other copyright owners. For more information on Open Research Online’s data policy on reuse of materials please consult the policies page. oro.open.ac.uk 1 Faculty of Biology. Dissertation for the Degree of Bachelor of Philosophy, Susan Brown North London Blood Transfusion Centre Colindale. Detailed follow-up of hepatitis B surface antigen positive blood donors. fMav 1992. ProQ uest Number: 27919423 All rights reserved INFORMATION TO ALL USERS The quality of this reproduction is dependent on the quality of the copy submitted. in the unlikely event that the author did not send a complete manuscript and there are missing pages, these will be noted. Also, if material had to be removed, a note will indicate the deletion. uest ProQuest 27919423 Published by ProQuest LLC (2020). Copyright of the Dissertation is held by the Author. Ail Rights Reserved. This work is protected against unauthorized copying under Title 17, United States Code Microform Edition © ProQuest LLC.
    [Show full text]
  • A Two-Site Immunoradiometric Assay for Serum Calcitonin Using Monoclonal Anti-Peptide Antibodies
    Henry Ford Hospital Medical Journal Volume 35 Number 2 Second International Workshop on Article 15 MEN-2 6-1987 A Two-Site Immunoradiometric Assay for Serum Calcitonin Using Monoclonal Anti-Peptide Antibodies Philippe Motte Malika Ait-Abdellah Pascal Vauzelle Paule Gardet Claude Bohuon See next page for additional authors Follow this and additional works at: https://scholarlycommons.henryford.com/hfhmedjournal Part of the Life Sciences Commons, Medical Specialties Commons, and the Public Health Commons Recommended Citation Motte, Philippe; Ait-Abdellah, Malika; Vauzelle, Pascal; Gardet, Paule; Bohuon, Claude; and Bellet, Dominique (1987) "A Two-Site Immunoradiometric Assay for Serum Calcitonin Using Monoclonal Anti- Peptide Antibodies," Henry Ford Hospital Medical Journal : Vol. 35 : No. 2 , 129-132. Available at: https://scholarlycommons.henryford.com/hfhmedjournal/vol35/iss2/15 This Article is brought to you for free and open access by Henry Ford Health System Scholarly Commons. It has been accepted for inclusion in Henry Ford Hospital Medical Journal by an authorized editor of Henry Ford Health System Scholarly Commons. A Two-Site Immunoradiometric Assay for Serum Calcitonin Using Monoclonal Anti-Peptide Antibodies Authors Philippe Motte, Malika Ait-Abdellah, Pascal Vauzelle, Paule Gardet, Claude Bohuon, and Dominique Bellet This article is available in Henry Ford Hospital Medical Journal: https://scholarlycommons.henryford.com/ hfhmedjournal/vol35/iss2/15 A TVo-Site Immunoradiometric Assay for Serum Calcitonin Using Monoclonal Anti-Peptide Antibodies Philippe Motte,* Malika Ait-Abdellah, Pascal Vauzelle, Paule Gardet, Claude Bohuon, and Dominique Beliet We have produced a library of monoclonal antibodies of various affinities by immunizing mice with synthetic calcitonin (CT) 1-32. These monoclonal antibodies defined two antigenic determinants on the molecule ofCT.
    [Show full text]
  • LECTIN-ENZYME BINDING ASSAYS Development of the Technique and Applications in Biochemistry and Medicine
    LECTIN-ENZYME BINDING ASSAYS Development of the technique and applications in biochemistry and medicine LECTINE-ENZYM BINDINGS ANALYSES Ontwikkeling van de techniek en enige toepassingen in de biochemie en de geneeskunde PROEFSCHRIFT Ter verkrijging van de graad van doctor aan de Erasmus Universiteit Rotterdam op gezag van de Rector Magnificus Prof. Dr. A.H.G. Rinnooy Kan en volgens besluit van het College van Dekanen. De openbare verdediging zal plaatsvinden op woensdag 18 januari 1989 om 15.45 uur door Jan Maurits Pekelharing geboren te Eindhoven 1989 DRUKKERU J.H. PASMANS B.V., 's-GRAVENHAGE PROMOTIECOMMISSIE Promotor: Prof. Dr. B. Leijnse Overige leden: Prof. Dr. E.H. Cooper, MD (Leeds) Prof. Dr. H.G. van Eijk Prof. Dr. J.BJ. Soons (Utrecht) Aan moeder Saar, voor het creatieve Aan vader Johan, voor het nuchtere CIP-DATA KONINKLIJKE BIBLIOTHEEK, DEN HAAG Pekelha~ing, Jan Maurits Lectin-enzyme binding assays : development of the technique and applications in biochemistry and medicine I Jan Maurits Pekelharing. - CS.l. : s.n.J C's-Gravenhage Pasmans). -Ill. Thesis Rotterdam. -With index, ref. -With summary in Dutch ISBN 90-9002656-8 SISO 546 UDC 547.96(043.3) Subject heading: lectin-enzyme binding assays. 5 CONTENTS Chapter 1: Lectins and the investigation of plasma protein glycosylation 1.1. Introduction 11 1.2. Lectins 11 1.2.1. History 11 1.2.2. Definitions 12 1.2.3. Occurrence 13 1.2.4. Specificity 13 1.2.5. Lectin methods 14 1.3. Protein glycosylation 15 1.3.1. Roles of glycosylation 17 1.3.2. Changes in plasma glycoprotein glycosylation 20 1.4.
    [Show full text]
  • Clinical Applications of Direct Antiglobulin Test
    Blood, Heart and Circulation Review Article ISSN: 2515-091X Clinical applications of direct antiglobulin test Jeong-Shi Lin1,2,3* 1Division of Transfusion Medicine, Department of Medicine, Taipei Veterans General Hospital, Taipei, Taiwan 2Division of Hematology, Department of Medicine, Taipei Veterans General Hospital, Taipei, Taiwan 3National Yang-Ming University School of Medicine, Taipei, Taiwan Abstract The direct antiglobulin test (DAT) is used to detect immunoglobulin and/or complement on the surface of red blood cells (RBCs). The DAT is valuable in the investigation of autoimmune hemolytic anemia, drug-induced immune hemolysis, hemolytic disease of newborn, hemolytic transfusion reactions, and passenger lymphocyte syndrome. There are several limitations of DAT, such as sensitivity, false positive and false negative. The patient's clinical history, diagnoses, and other laboratory test results should also take into consideration for DAT interpretation. Introduction Gel microcolumn DATs were more sensitive than tube agglutination and affinity microcolumn DATs [12]. Gel microcolumn DAT is a The direct antiglobulin test (DAT), also known as ' direct Coombs’ better alternative to conventional tube DAT for detecting red cell test', was found more widespread notoriety after been described in 1945 bound antibodies in various clinical conditions [13]. The sensitive gel by Cambridge immunologist Robin Coombs. The DAT is used to detect technology has enabled the hematologist not only to diagnose some immunoglobulin, complement, or both on the surface of red blood AIHA patients with negative DAT, but also to characterize red cell cells (RBCs). The indirect antiglobulin test (IAT) is used to detect red bound autoantibodies with regard to their class, subclass and titer in a cell antibodies in patient serum.
    [Show full text]
  • Serology of Paracoccidioidomycosis
    View metadata, citation and similar papers at core.ac.uk brought to you by CORE provided by Repositório Institucional UNIFESP Mycopathologia (2008) 165:289–302 DOI 10.1007/s11046-007-9060-5 Serology of paracoccidioidomycosis Zoilo Pires de Camargo Received: 10 July 2007 / Accepted: 30 August 2007 Ó Springer Science+Business Media B.V. 2007 Abstract This review provides the background for Keywords Antigens Á Mycosis Á understanding the role of a battery of diagnostic Paracoccidioidomycosis Á Paracoccidioides methods in paracoccidioidomycosis (PCM). This brasiliensis Á Serology systemic mycosis is a disease endemic in many regions of Latin America, with sporadic cases also occurring throughout the world (mycosis of importa- Introduction tion). Although excellent laboratory methods for diagnosis are available, there are deficiencies that The fungus Paracoccidioides brasiliensis is must be met by continued research. Understanding ensconced in the nature in Latin America, but its the uses and limitations of a battery of laboratory exact niche is not known. The expanding human and methods is essential to diagnose PCM. Clinicians and non-human populations of endemic areas provide a laboratory directors must be familiar with the uses continuing supply of individuals who are susceptible and limitations of a battery of serologic and myco- to infection with P. brasiliensis. Paracoccidioidomy- logical tests to accurately diagnose of PCM. cosis (PCM) is caused by the inhalation of conidia Antibody and antigen detections are valuable found in nature and under adverse nutritional condi- adjuncts to histopathology and culture. More tions, the fungus sporulates forming conidia which recently, the gp43 and gp70 antigen detection assay are less than 5 lm in diameter and can easily reach have improved the methodology of diagnosis of this the alveoli when inhaled, and a lung-lymph node mycosis, which improves reproducibility and facili- primary complex develops.
    [Show full text]
  • Cross-Talk Between Gonadotropin- Releasing
    CROSS-TALK BETWEEN GONADOTROPIN- RELEASING HORMONES AND PROGESTERONE RECEPTOR IN NEUROENDOCRINE CELLS by BEUM-SOO AN D.V.M., Chung-buk National University, 2000 M.Sc, Chung-buk National University, 2002 A THESIS SUBMITTED IN PARTIAL FULFILLMENT OF THE REQUIREMENTS FOR THE DEGREE OF DOCTOR OF PHILOSOPHY in THE FACULTY OF GRADUATE STUDIES (Reproductive & Developmental Sciences) THE UNIVERSITY OF BRITISH COLUMBIA March 2007 © Beum-soo AN, 2007 ABSTRACT Hypothalamic gonadotropin-releasing hormone (GnRH) is a decapeptide that plays a pivotal role in mammalian reproduction. It is hypothesized that progesterone (P4) may regulate GnRH I, GnRH II (a second form of GnRH) and GnRH I receptor (GnRH I R) at the transcriptional level. Alternatively, GnRHs may stimulate transactivation of the progesterone receptor (PR), thereby, modulating gonadotropin subunit gene expression. Treatment of human neuronal cells with P4 suppressed GnRH I R promoter activity. This P4-stimulated inhibition was enhanced when PR A was over-expressed. With respect to the two GnRHs, P4 increased GnRH I mRNA levels, but did not significantly affect GnRH II gene expression. Regulation of gonadotropin production involves interplay between steroids and neuro• peptides, thus we have examined the effects of GnRHs on PR activation in pituitary cells. Treatment with GnRHs increased a progesterone response element (PRE)-luciferase reporter gene activity. PR was phosphorylated at Ser294 and translocated into nucleus after GnRH treatment in the absence of P4. Interactions between the PR and several coactivators were examined, and treatment with GnRHs specifically induced PR: Steroid Receptor Coactivator-3 (SRC-3) interaction. In chromatin immunoprecipitation assays, recruitment of PR and SRC-3 to the PRE reporter gene was also increased by GnRHs.
    [Show full text]