logging in or signing up serology Amateur Download Post to : URL : Related Presentations : Share Add to Flag Embed Email Send to Blogs and Networks Add to Channel Uploaded from authorPOINT Insert YouTube videos in PowerPont slides with aS Desktop Copy embed code: (To copy code, click on the text box) Embed: URL: Thumbnail: WordPress Embed Customize Embed The presentation is successfully added In Your Favorites. Views: 4623 Category: Entertainment License: All Rights Reserved Like it (6) Dislike it (0) Added: June 19, 2007 This Presentation is Public Favorites: 2 Presentation Description No description available. Comments Posting comment... By: c2di (15 month(s) ago) i like this presentation. please forward to my email : patelsp21@gmail.com pleaseeeeeeeeeeeeeeeeeeeeeeeeee Saving..... Post Reply Close Saving..... Edit Comment Close By: iAMkimie (18 month(s) ago) Great presentation. Can I have a copy please? Please forward to my email: khimmie02@yahoo.com Saving..... Post Reply Close Saving..... Edit Comment Close By: ascaris99 (31 month(s) ago) may i please download this as powerpoint file? i wish to use some of your data in my report. thank you very much! jun Saving..... Post Reply Close Saving..... Edit Comment Close Premium member Presentation Transcript Slide1: Sean K. Bauman, Ph.D. V.P. Operations Immuno-Mycologics, Inc. Serodiagnosis of Mycoses Slide2: Outline Why use serology? Mechanisms of different tests Serology of specific mycoses Slide3: Why Use Serology For Case Finding? Antigens and antibodies are easier to detect than finding the organism directly. Antibodies are produced in large quantities and circulate in the blood even though they are not usually protective against mycotic agents. Soluble antigens frequently enter the circulation of the host and are sometimes excreted in the urine. They are produced in discrete infected lesions. Culture is relatively Insensitive due to the low concentration of the agents in tissues. Multiple cultures are usually required. Slide4: Immunobiology, 4th Edition Immunodiffusion Slide5: Immunobiology, 4th Edition Immunodiffusion Concentration Slide6: Immunobiology, 4th Edition Immunodiffusion Concentration Slide7: Kuby Immunology, 4th Edition Immunodiffusion Slide8: Kuby Immunology, 4th Edition Immunodiffusion Slide9: Kuby Immunology, 4th Edition Immunodiffusion Slide10: Immunodiffusion Types of Immunodiffusion Reactions: Identity Partial Identity Non-Identity Slide11: Immunodiffusion Identity Adapted from Kuby Immunology, 4th Edition Slide12: Immunodiffusion Partial Identity Adapted from Kuby Immunology, 4th Edition Slide13: Immunodiffusion Non-Identity Adapted from Kuby Immunology, 4th Edition Slide14: Immunodiffusion Slide15: Qualitative vs Quantitative Immunodiffusion Positive Undiluted Positive @ 1:4 Identity andamp; Partial Identity bands are considered POSITIVE Slide16: Complement Fixation Day 1 Day 2 Slide17: Complement Fixation Slide18: Latex Agglutination Antigen Test Slide19: Latex Agglutination Slide20: Sandwich ELISA Adapted from Kuby Immunology, 4th Edition Enzyme-Linked Immunosorbent Assay Antigen Test Slide21: Sandwich ELISA Adapted from Kuby Immunology, 4th Edition Enzyme-Linked Immunosorbent Assay Add Specimen Antigen Test Slide22: Sandwich ELISA Adapted from Kuby Immunology, 4th Edition Enzyme-Linked Immunosorbent Assay Add Specimen Antigen Test Slide23: Sandwich ELISA Adapted from Kuby Immunology, 4th Edition Enzyme-Linked Immunosorbent Assay Add Specimen Containing Ag Antigen Test Slide24: Indirect ELISA Adapted from Kuby Immunology, 4th Edition Enzyme-Linked Immunosorbent Assay Antibody Test Slide25: Adapted from Kuby Immunology, 4th Edition Enzyme-Linked Immunosorbent Assay (Specimen) Indirect ELISA Antibody Test Slide26: Adapted from Kuby Immunology, 4th Edition Enzyme-Linked Immunosorbent Assay (Specimen) Indirect ELISA Antibody Test Slide27: Adapted from Kuby Immunology, 4th Edition Enzyme-Linked Immunosorbent Assay (Specimen) Indirect ELISA Antibody Test Slide28: Mycotic Diseases Aspergillosis Blastomycosis Candidiasis Coccidioidomycosis Cryptococcosis Histoplasmosis Paracoccidioidomycosis Organism, Disease, Diagnosis Slide29: Aspergillosis The Organism Slide30: Infection initiates by inhalation of Aspergillus spores Aspergillosis The Disease Allergic Bronchopulmonary Aspergillosis (ABPA) Pulmonary Aspergilloma (fungus ball) Invasive Aspergillosis (IA) (usually immunocompromised) Forms of the Disease Slide31: Aspergillosis Antibody Test 1 band indicates current/recent Aspergillus infection Immunodiffusion Species # Bands Sensitivity Specificity A. fumigatus 2 90% 99.6% A. flavus 1 100% 100% A. niger 1 100% 100% A. terreus 1 n.d. n.d. 3 bands is indicative of Aspergilloma andamp;/or IA Sabouraudia (1982) 20:63 Diagnosis Slide32: Aspergillosis Antigen Tests Latex Agglutination ELISA Sensitivity Specificity Pastorex LA 31-95% 91-95% Platelia ELISA 60-93% 82-99% Clin Micro Rev (2002) 15:465 Diagnosis Slide33: Blastomycosis Etiologic agent: Blastomyces dermatitidis The Organism Slide34: Blastomycosis Infection initiates by inhalation of the organism The Disease Slide35: Blastomycosis Antibody Test ‘A’ antigen (a.k.a. WI-1) 25-amino acid tandem repeat is immunodominant epitope Precipitin band indicates current/recent B. dermatitidis infection 120-kDa glycoprotein Immunodiffusion Diagnosis Complement Fixation Slide36: Candidiasis Primary etiologic agent: Candida albicans C. albicans is part of the normal human flora The Organism Slide37: Candidiasis Immunocompromised states susceptible to Candidiasis Neutropenia Corticosteroids Diabetes mellitus HIV infection Cancer Infection types: The Disease Vaginal Antibiotic therapy Slide38: Candidiasis Antibody Test Systemic candidiasis should be suspected when 1 band indicates current/recent infection Seroconversion (i.e. negative patient becomes positive) Sensitivity is less in immunodeficient individuals Immunodiffusion Diagnosis Slide39: Candidiasis Antigen/Metabolite Tests Latex Agglutination ELISA b-Glucan Cand-Tec assay LA 77% 88% Clin Micro Rev (2002) 15:465 Sensitivity Specificity Pastorex LA 25-28% 100% Platelia ELISA 42% 98% b-Glucan Limulus 84% 88% Diagnosis Slide40: Coccidioidomycosis The Organism Slide41: Coccidioidomycosis The Disease Slide42: Coccidioidomycosis Antibody Test Sensitivity 51% Specificity 89% J. Clin. Micro. (1995) 33:940 Diagnosis Sensitivity 89% Specificity 89% Sensitivity 94% Specificity 92% Latex Agglutination Complement Fixation ELISA Slide43: Heat labile antigen Predominately IgG reactivity Heat stable antigen (60°C, 30 min) 3-O-methylmannose (3-O-MM) is immunodominant epitope IDTP 120-kDa glycoprotein (BGL2) Predominantly IgM reactivity in early primary cases Closest to the antigen well Immunodiffusion Coccidioidomycosis Antibody Test Diagnosis Slide44: Cryptococcosis The Organism Slide45: Cryptococcosis The Disease Immunocompromised (e.g. AIDS) are most susceptible Adapted from Clinical microbiology made ridiculously simple by M. Gladwin and B. Trattler Slide46: Cryptococcosis Antigen Tests Sensitivity 100% Specificity 100% Mycoses (1993) 36:31 Sensitivity 85% Specificity 97% Diagnosis Latex Agglutination ELISA Slide47: Histoplasmosis The Organism Slide48: Histoplasmosis The Disease Infection initiates by inhalation of the organism Intracellular proliferation Slide49: Histoplasmosis Antibody Test M Antigen 81-kDa glycoprotein (catalase) M precipitins first to appear in acute disease H Antigen 93-kDa glycoprotein (b-glucosidase) Closest to the antigen well Closest to the serum well Immunodiffusion J. Clin. Micro. (1976) 2:77 Diagnosis Slide50: Complement Fixation Sensitivity 94% Specificity 95% J. Clin. Micro. (1976) 2:77 Histoplasmosis Antibody Test Diagnosis Slide51: Histoplasmosis Antigen Tests EIA/RIA Urine RIA 89% 90% Sensitivity Specificity Serum RIA 82% 77% Serum EIA 68% 81% Urine EIA 89% 90% Clin Micro Rev (2002) 15:465 Diagnosis Slide52: Paracoccidioidomycosis The Organism Slide53: The Disease Paracoccidioidomycosis Slide54: Paracoccidioidomycosis Antibody Test Sensitivity = andgt;90% * Manual of Clinical Laboratory Immunology, 6th Edition Up to 3 precipitin bands are observed Most prevalent and longest lasting of the major precipitins Immunodiffusion Diagnosis Slide55: Why Use Serology For Case Finding? Antigens and antibodies are easier to detect than finding the organism directly. Antibodies are produced in large quantities and circulate in the blood even though they are not usually protective against mycotic agents. Soluble antigens frequently enter the circulation of the host and are sometimes excreted in the urine. They are produced in discrete infected lesions. Culture is relatively Insensitive due to the low concentration of the agents in tissues. Multiple cultures are usually required. Slide56: Do one thing, do it well! The Fungal Diagnostic Specialists www.immy.com You do not have the permission to view this presentation. In order to view it, please contact the author of the presentation.
serology Amateur Download Post to : URL : Related Presentations : Share Add to Flag Embed Email Send to Blogs and Networks Add to Channel Uploaded from authorPOINT Insert YouTube videos in PowerPont slides with aS Desktop Copy embed code: (To copy code, click on the text box) Embed: URL: Thumbnail: WordPress Embed Customize Embed The presentation is successfully added In Your Favorites. Views: 4623 Category: Entertainment License: All Rights Reserved Like it (6) Dislike it (0) Added: June 19, 2007 This Presentation is Public Favorites: 2 Presentation Description No description available. Comments Posting comment... By: c2di (15 month(s) ago) i like this presentation. please forward to my email : patelsp21@gmail.com pleaseeeeeeeeeeeeeeeeeeeeeeeeee Saving..... Post Reply Close Saving..... Edit Comment Close By: iAMkimie (18 month(s) ago) Great presentation. Can I have a copy please? Please forward to my email: khimmie02@yahoo.com Saving..... Post Reply Close Saving..... Edit Comment Close By: ascaris99 (31 month(s) ago) may i please download this as powerpoint file? i wish to use some of your data in my report. thank you very much! jun Saving..... Post Reply Close Saving..... Edit Comment Close Premium member Presentation Transcript Slide1: Sean K. Bauman, Ph.D. V.P. Operations Immuno-Mycologics, Inc. Serodiagnosis of Mycoses Slide2: Outline Why use serology? Mechanisms of different tests Serology of specific mycoses Slide3: Why Use Serology For Case Finding? Antigens and antibodies are easier to detect than finding the organism directly. Antibodies are produced in large quantities and circulate in the blood even though they are not usually protective against mycotic agents. Soluble antigens frequently enter the circulation of the host and are sometimes excreted in the urine. They are produced in discrete infected lesions. Culture is relatively Insensitive due to the low concentration of the agents in tissues. Multiple cultures are usually required. Slide4: Immunobiology, 4th Edition Immunodiffusion Slide5: Immunobiology, 4th Edition Immunodiffusion Concentration Slide6: Immunobiology, 4th Edition Immunodiffusion Concentration Slide7: Kuby Immunology, 4th Edition Immunodiffusion Slide8: Kuby Immunology, 4th Edition Immunodiffusion Slide9: Kuby Immunology, 4th Edition Immunodiffusion Slide10: Immunodiffusion Types of Immunodiffusion Reactions: Identity Partial Identity Non-Identity Slide11: Immunodiffusion Identity Adapted from Kuby Immunology, 4th Edition Slide12: Immunodiffusion Partial Identity Adapted from Kuby Immunology, 4th Edition Slide13: Immunodiffusion Non-Identity Adapted from Kuby Immunology, 4th Edition Slide14: Immunodiffusion Slide15: Qualitative vs Quantitative Immunodiffusion Positive Undiluted Positive @ 1:4 Identity andamp; Partial Identity bands are considered POSITIVE Slide16: Complement Fixation Day 1 Day 2 Slide17: Complement Fixation Slide18: Latex Agglutination Antigen Test Slide19: Latex Agglutination Slide20: Sandwich ELISA Adapted from Kuby Immunology, 4th Edition Enzyme-Linked Immunosorbent Assay Antigen Test Slide21: Sandwich ELISA Adapted from Kuby Immunology, 4th Edition Enzyme-Linked Immunosorbent Assay Add Specimen Antigen Test Slide22: Sandwich ELISA Adapted from Kuby Immunology, 4th Edition Enzyme-Linked Immunosorbent Assay Add Specimen Antigen Test Slide23: Sandwich ELISA Adapted from Kuby Immunology, 4th Edition Enzyme-Linked Immunosorbent Assay Add Specimen Containing Ag Antigen Test Slide24: Indirect ELISA Adapted from Kuby Immunology, 4th Edition Enzyme-Linked Immunosorbent Assay Antibody Test Slide25: Adapted from Kuby Immunology, 4th Edition Enzyme-Linked Immunosorbent Assay (Specimen) Indirect ELISA Antibody Test Slide26: Adapted from Kuby Immunology, 4th Edition Enzyme-Linked Immunosorbent Assay (Specimen) Indirect ELISA Antibody Test Slide27: Adapted from Kuby Immunology, 4th Edition Enzyme-Linked Immunosorbent Assay (Specimen) Indirect ELISA Antibody Test Slide28: Mycotic Diseases Aspergillosis Blastomycosis Candidiasis Coccidioidomycosis Cryptococcosis Histoplasmosis Paracoccidioidomycosis Organism, Disease, Diagnosis Slide29: Aspergillosis The Organism Slide30: Infection initiates by inhalation of Aspergillus spores Aspergillosis The Disease Allergic Bronchopulmonary Aspergillosis (ABPA) Pulmonary Aspergilloma (fungus ball) Invasive Aspergillosis (IA) (usually immunocompromised) Forms of the Disease Slide31: Aspergillosis Antibody Test 1 band indicates current/recent Aspergillus infection Immunodiffusion Species # Bands Sensitivity Specificity A. fumigatus 2 90% 99.6% A. flavus 1 100% 100% A. niger 1 100% 100% A. terreus 1 n.d. n.d. 3 bands is indicative of Aspergilloma andamp;/or IA Sabouraudia (1982) 20:63 Diagnosis Slide32: Aspergillosis Antigen Tests Latex Agglutination ELISA Sensitivity Specificity Pastorex LA 31-95% 91-95% Platelia ELISA 60-93% 82-99% Clin Micro Rev (2002) 15:465 Diagnosis Slide33: Blastomycosis Etiologic agent: Blastomyces dermatitidis The Organism Slide34: Blastomycosis Infection initiates by inhalation of the organism The Disease Slide35: Blastomycosis Antibody Test ‘A’ antigen (a.k.a. WI-1) 25-amino acid tandem repeat is immunodominant epitope Precipitin band indicates current/recent B. dermatitidis infection 120-kDa glycoprotein Immunodiffusion Diagnosis Complement Fixation Slide36: Candidiasis Primary etiologic agent: Candida albicans C. albicans is part of the normal human flora The Organism Slide37: Candidiasis Immunocompromised states susceptible to Candidiasis Neutropenia Corticosteroids Diabetes mellitus HIV infection Cancer Infection types: The Disease Vaginal Antibiotic therapy Slide38: Candidiasis Antibody Test Systemic candidiasis should be suspected when 1 band indicates current/recent infection Seroconversion (i.e. negative patient becomes positive) Sensitivity is less in immunodeficient individuals Immunodiffusion Diagnosis Slide39: Candidiasis Antigen/Metabolite Tests Latex Agglutination ELISA b-Glucan Cand-Tec assay LA 77% 88% Clin Micro Rev (2002) 15:465 Sensitivity Specificity Pastorex LA 25-28% 100% Platelia ELISA 42% 98% b-Glucan Limulus 84% 88% Diagnosis Slide40: Coccidioidomycosis The Organism Slide41: Coccidioidomycosis The Disease Slide42: Coccidioidomycosis Antibody Test Sensitivity 51% Specificity 89% J. Clin. Micro. (1995) 33:940 Diagnosis Sensitivity 89% Specificity 89% Sensitivity 94% Specificity 92% Latex Agglutination Complement Fixation ELISA Slide43: Heat labile antigen Predominately IgG reactivity Heat stable antigen (60°C, 30 min) 3-O-methylmannose (3-O-MM) is immunodominant epitope IDTP 120-kDa glycoprotein (BGL2) Predominantly IgM reactivity in early primary cases Closest to the antigen well Immunodiffusion Coccidioidomycosis Antibody Test Diagnosis Slide44: Cryptococcosis The Organism Slide45: Cryptococcosis The Disease Immunocompromised (e.g. AIDS) are most susceptible Adapted from Clinical microbiology made ridiculously simple by M. Gladwin and B. Trattler Slide46: Cryptococcosis Antigen Tests Sensitivity 100% Specificity 100% Mycoses (1993) 36:31 Sensitivity 85% Specificity 97% Diagnosis Latex Agglutination ELISA Slide47: Histoplasmosis The Organism Slide48: Histoplasmosis The Disease Infection initiates by inhalation of the organism Intracellular proliferation Slide49: Histoplasmosis Antibody Test M Antigen 81-kDa glycoprotein (catalase) M precipitins first to appear in acute disease H Antigen 93-kDa glycoprotein (b-glucosidase) Closest to the antigen well Closest to the serum well Immunodiffusion J. Clin. Micro. (1976) 2:77 Diagnosis Slide50: Complement Fixation Sensitivity 94% Specificity 95% J. Clin. Micro. (1976) 2:77 Histoplasmosis Antibody Test Diagnosis Slide51: Histoplasmosis Antigen Tests EIA/RIA Urine RIA 89% 90% Sensitivity Specificity Serum RIA 82% 77% Serum EIA 68% 81% Urine EIA 89% 90% Clin Micro Rev (2002) 15:465 Diagnosis Slide52: Paracoccidioidomycosis The Organism Slide53: The Disease Paracoccidioidomycosis Slide54: Paracoccidioidomycosis Antibody Test Sensitivity = andgt;90% * Manual of Clinical Laboratory Immunology, 6th Edition Up to 3 precipitin bands are observed Most prevalent and longest lasting of the major precipitins Immunodiffusion Diagnosis Slide55: Why Use Serology For Case Finding? Antigens and antibodies are easier to detect than finding the organism directly. Antibodies are produced in large quantities and circulate in the blood even though they are not usually protective against mycotic agents. Soluble antigens frequently enter the circulation of the host and are sometimes excreted in the urine. They are produced in discrete infected lesions. Culture is relatively Insensitive due to the low concentration of the agents in tissues. Multiple cultures are usually required. Slide56: Do one thing, do it well! The Fungal Diagnostic Specialists www.immy.com