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Histopathology and Laboratory Features of Sexually Transmitted Diseases
Histopath & Labs for STIs Endo, Energy and Repro 2017-2018 HISTOPATHOLOGY AND LABORATORY FEATURES OF SEXUALLY TRANSMITTED DISEASES Dominck Cavuoti, D.O. Phone: 469-419-3412 Email: [email protected] LEARNING OBJECTIVES: • Identify the etiologic agents causing pelvic inflammatory disease and the pathologic changes they produce. • Discuss the characteristic clinical and pathologic findings caused by herpes simplex virus (HSV) infections: a. fever blisters b. genital herpes simplex virus infection c. disseminated neonatal HSV • Describe the pathologic changes produced by Treponema pallidum. • Describe the clinical features and pathologic changes produced by Chlamydia trachomatisand Neisseria gonorrhoeae • Describe the clinical and laboratory features of vaginal infections including: Trichomonas, Candida, and bacterial vaginosis. • Describe the clinical and laboratory features of ectoparasite infections PURPOSE OF THE LECTURE: 1. To describe the various agents of sexually transmitted diseases and their disease manifestations 2. To describe the pathologic features associated with STDs 3. To introduce some of the laboratory aspects of STDs TERMS INTRODUCED IN LECTURE: Condyloma lata Disseminated gonococcal infection Gummatous syphilis Lymphogranuloma venereum Pelvic inflammatory disease Rapid Plasma Reagin (RPR) Salpingitis Syphilis/endarteritis obliterans Venereal Disease Research Laboratory (VDRL) Treponema pallidum particle agglutination (TPPA) Histopath & Labs for STIs Endo, Energy and Repro 2017-2018 MAJOR CONCEPTS EMPHASIZED IN LECTURE I. Syphilis (Will be covered by Dr. Norgard in later lecture). II. Gonorrhea A. Causative agent: Neisseria gonorrhoeae, a Gram negative diplococcus. Humans are the only natural reservoir. Infection is acquired via direct contact with the mucosa of an infected person. The incubation period averages 2-5 days with a range of 1-14 days. -
Francisella Tularensis 6/06 Tularemia Is a Commonly Acquired Laboratory Colony Morphology Infection; All Work on Suspect F
Francisella tularensis 6/06 Tularemia is a commonly acquired laboratory Colony Morphology infection; all work on suspect F. tularensis cultures .Aerobic, fastidious, requires cysteine for growth should be performed at minimum under BSL2 .Grows poorly on Blood Agar (BA) conditions with BSL3 practices. .Chocolate Agar (CA): tiny, grey-white, opaque A colonies, 1-2 mm ≥48hr B .Cysteine Heart Agar (CHA): greenish-blue colonies, 2-4 mm ≥48h .Colonies are butyrous and smooth Gram Stain .Tiny, 0.2–0.7 μm pleomorphic, poorly stained gram-negative coccobacilli .Mostly single cells Growth on BA (A) 48 h, (B) 72 h Biochemical/Test Reactions .Oxidase: Negative A B .Catalase: Weak positive .Urease: Negative Additional Information .Can be misidentified as: Haemophilus influenzae, Actinobacillus spp. by automated ID systems .Infective Dose: 10 colony forming units Biosafety Level 3 agent (once Francisella tularensis is . Growth on CA (A) 48 h, (B) 72 h suspected, work should only be done in a certified Class II Biosafety Cabinet) .Transmission: Inhalation, insect bite, contact with tissues or bodily fluids of infected animals .Contagious: No Acceptable Specimen Types .Tissue biopsy .Whole blood: 5-10 ml blood in EDTA, and/or Inoculated blood culture bottle Swab of lesion in transport media . Gram stain Sentinel Laboratory Rule-Out of Francisella tularensis Oxidase Little to no growth on BA >48 h Small, grey-white opaque colonies on CA after ≥48 h at 35/37ºC Positive Weak Negative Positive Catalase Tiny, pleomorphic, faintly stained, gram-negative coccobacilli (red, round, and random) Perform all additional work in a certified Class II Positive Biosafety Cabinet Weak Negative Positive *Oxidase: Negative Urease *Catalase: Weak positive *Urease: Negative *Oxidase, Catalase, and Urease: Appearances of test results are not agent-specific. -
Biofire Blood Culture Identification System (BCID) Fact Sheet
BioFire Blood Culture Identification System (BCID) Fact Sheet What is BioFire BioFire BCID is a multiplex polymerase chain reaction (PCR) test designed to BCID? identify 24 different microorganism targets and three antibiotic resistance genes from positive blood culture bottles. What is the purpose The purpose of BCID is to rapidly identify common microorganisms and of BCID? antibiotic resistance genes from positive blood cultures so that antimicrobial therapy can be quickly optimized by the physician and the antibiotic stewardship pharmacist. It is anticipated that this will result in improved patient outcomes, decreased length of stay, improved antibiotic stewardship, and decreased costs. When will BCID be BCID is performed on all initially positive blood cultures after the gram stain is routinely performed and reported. performed? When will BCID not For blood cultures on the same patient that subsequently become positive with be routinely a microorganism showing the same morphology as the initial positive blood performed? culture, BCID will not be performed. BCID will not be performed on positive blood cultures with gram positive bacilli unless Listeria is suspected. BCID will not be performed on blood culture bottles > 8 hours after becoming positive. BCID will not be performed between 10PM-7AM on weekdays and 2PM-7AM on weekends. BCID will not be performed for clinics that have specifically opted out of testing. How soon will BCID After the blood culture becomes positive and the gram stain is performed and results be available? reported, the bottle will be sent to the core Microbiology lab by routine courier. BCID testing will then be performed. It is anticipated that total turnaround time will generally be 2-3 hours after the gram stain is reported. -
Interpreting and Acting on Positive Blood Cultures Trevor Van Schooneveld, MD 1/18/18 Objectives
Stewardship Interventions: Interpreting and Acting on Positive Blood Cultures Trevor Van Schooneveld, MD 1/18/18 Objectives • Interpret the results of blood cultures including gram stains and rapid pathogen diagnostic tests • Make recommendations regarding antimicrobial therapy based on interpretation of blood culture data Early Initiation of Active Therapy is Essential Predicted hospital mortality and 95% CIs for time to first antibiotic administration Surviving Sepsis Guidelines (N=28,150 severe sepsis, septic shock patients) • Administer IV antimicrobials within one hour of presentation (strong) • Initiate empiric, broad-spectrum therapy with one or more agents to cover all likely pathogens (strong) Ferrer R, et al. Crit Care Med. 2014;42:1749-55. Rhodes A, et al. Crit Care Med. 2017;45:486-552. De-escalation Also is Important Surviving Sepsis Guidelines • Narrow empiric antibiotics once pathogen identified and/or clinical improvement De-escalation Benefit • De-escalation in severe sepsis, septic shock (N=712) • Mortality OR 0.54 (95% CI 0.33-0.89, P=.016) • De-escalation in community-onset gram-negative bacteremia (N=189) • Mortality OR 0.37 (0.14-0.96, P=.04) Garnarcho-Montero J, et al. Intensive Care Med. 2014;40:32-40. Lee C, et al. Diag Micro Infect Dis. 2015;82:158-64. Issues with Treatment of Sepsis/Bacteremia Under-treatment • May die (mortality) • May not get better as quickly (LOS, cost) • May develop complications (LOS, cost) Overtreatment • May develop toxicities (cost, LOS) • May develop C. difficile (cost, LOS, readmission) -
BLOOD CULTURE MEDIA Principle: Specimen: Reagent Preparation Storage: Procedure
BLOOD CULTURE MEDIA For In-Vitro and professional use only Store at (2° to 8°C) Blood cultures are used to detect the presence of bacteria or fungi in the blood, to identify the type present, and to guide treatment. Testing is used to identify a blood infection (septicemia) that can lead to sepsis, a serious and life-threatening complication. Individuals with a suspected blood infection are often treated in intensive care units, so testing is often done in a hospital setting. A bacterial infection in the blood called bacteremia. It can be serious because the blood can spread the bacteria to any part of the body. Blood infections most often occur with other serious infections such as those affecting the lungs, kidneys, bowel, gallbladder, or heart valves. Blood infections may also develop when the immune system is weak in infants and older adults, from disease (such as cancer or AIDS) or from medicines (such as corticosteroids or chemotherapy) that change the ability of your body to fight infections (immunity). Principle: The vials containing 25 ml or 50 ml of brain heart infusion, yeast extract, SPS and other stabilizers. The Media is used for yeast, aerobic and anaerobic organisms in blood. The principle of the this test is that each type of organisms need a certain time to grow and multiply. Specimen: Blood. Reagent preparation The vials are ready to use. Storage: Store reagent from (2 - 8 oC). Procedure: 1. Bottles of Brain Heart Infusion which are not used the same day as sterilized should be placed in a boiling water bath for several minutes to remove absorbed oxygen , and cooled rapidly without shaking , just before use. -
Section 13. Laboratory Considerations
Section 13. Laboratory Considerations Table of Contents 13.1 Overview and General Guidance 13.2 Specimen Labeling 13.3 Procedures for Specimens That Cannot be Evaluated 13.4 Use of LDMS 13.5 Documentation 13.6 Urine Testing 13.6.1 Specimen Collection 13.6.2 Pregnancy Testing 13.6.3 Chlamydia and Gonorrhea Testing 13.6.4 Urine Culture 13.7 Blood Testing 13.7.1 Specimen Collection and Initial Processing 13.7.2 HIV Testing 13.7.3 Syphilis Testing 13.7.4 Hematology Testing 13.7.5 Serum Chemistries 13.7.6 Plasma Storage 13.7.7 CD4+ T Cell Count 13.7.8 HIV RNA PCR 13.7.9 HIV DNA PCR 13.8 Testing of Vaginal and Cervical Specimens 13.8.1 Vaginal pH 13.8.2 Wet Mount for Candidiasis and BV 13.8.3 Rapid Test for Trichomoniasis 13.8.4 Vaginal Gram Stain 13.8.5 Papanicolaou (Pap) Test 13.8.6 Self-Administered Vaginal Swabs for PK and biomarker testing 13.8.7 Endocervical Swabs for Biomarker Analysis 13.8.8 Intra-Vaginal Ring Storage 13.8.9 Herpes Lesion Testing Table 13-1 Volume Guide for Plasma Storage Appendix 13-1 Overview of Laboratory Testing Locations, Specimens, and Methods for MTN-020 Appendix 13-2 MTN-020 Lab Specimen Processing Guidelines Appendix 13-3 LDMS Specimen Management Guide to Logging in MTN-020 Specimens Appendix 13-4 MTN-020 HIV Testing Algorithms Appendix 13-5 MTN Network Lab HIV Query Form Appendix 13-6 LDMS Tracking Sheets This section contains information on the laboratory procedures performed in MTN-020. -
Use of the Diagnostic Bacteriology Laboratory: a Practical Review for the Clinician
148 Postgrad Med J 2001;77:148–156 REVIEWS Postgrad Med J: first published as 10.1136/pmj.77.905.148 on 1 March 2001. Downloaded from Use of the diagnostic bacteriology laboratory: a practical review for the clinician W J Steinbach, A K Shetty Lucile Salter Packard Children’s Hospital at EVective utilisation and understanding of the Stanford, Stanford Box 1: Gram stain technique University School of clinical bacteriology laboratory can greatly aid Medicine, 725 Welch in the diagnosis of infectious diseases. Al- (1) Air dry specimen and fix with Road, Palo Alto, though described more than a century ago, the methanol or heat. California, USA 94304, Gram stain remains the most frequently used (2) Add crystal violet stain. USA rapid diagnostic test, and in conjunction with W J Steinbach various biochemical tests is the cornerstone of (3) Rinse with water to wash unbound A K Shetty the clinical laboratory. First described by Dan- dye, add mordant (for example, iodine: 12 potassium iodide). Correspondence to: ish pathologist Christian Gram in 1884 and Dr Steinbach later slightly modified, the Gram stain easily (4) After waiting 30–60 seconds, rinse with [email protected] divides bacteria into two groups, Gram positive water. Submitted 27 March 2000 and Gram negative, on the basis of their cell (5) Add decolorising solvent (ethanol or Accepted 5 June 2000 wall and cell membrane permeability to acetone) to remove unbound dye. Growth on artificial medium Obligate intracellular (6) Counterstain with safranin. Chlamydia Legionella Gram positive bacteria stain blue Coxiella Ehrlichia Rickettsia (retained crystal violet). -
Gram Stain Workshop for the Laboratory Generalist
Gram Stain Workshop for the Laboratory Generalist Karen Stiles, SM(ASCP)CM State Training Coordinator Assistant Chemical Terrorism Coordinator Nebraska Public Health Laboratory 402-559-3590 [email protected] 1 GRAM STAIN OBJECTIVES: Upon completion, the participant will be able to: 1. Explain the principle of the Gram stain procedure, including what elements can affect staining results 2. Correlate the most common pathogens with positive Gram stains from blood cultures and direct specimen sterile body fluid smears 3. Perform and interpret Grams stains 2 Purpose of Gram Stain Classify bacteria based on form, size, cellular morphology, Gram reaction Assess quality of specimen Identify specific infectious agent from morphology and Gram reaction Correlation with culture growth Correlation with culture-independent methodologist Guide presumptive antibiotic therapy 3 Principle of Gram Stain Cell wall composition Gram positive – think peptidoglycan layer with teichoic acid Gram negative – high in lipid content Basic premise Crystal Violet – all cells take up primary stain Gram’s iodine – mordant to form complex Decolorizer – mixture of acetone and alcohol Dehydrate lipids in Gram negative cell walls, wash out complex Gram positive cells resistant, retain stain complex Safranin - counterstain 4 Gram negative cells take up counterstain Preparation of Samples Specimen Type Preparation CSF/sterile body fluids Cyto/Centrifuge Blood Culture Broth Drop to slide Tissue Touch prep Tissue homogenate Drop to slide Swabbed material Roll -
Syphilis Diagnosis: Three Cases with Increasing Treponemal Test Result After Therapy
Acta Derm Venereol 2014; 94: 323–324 SHORT COMMUNICATION Syphilis Diagnosis: Three Cases with Increasing Treponemal Test Result after Therapy Henrik O. Larsson1*, Annika Johnsson2 and Anders Bredberg1 Departments of 1Medical Microbiology, and 2Dermatology and Venereology, Skane University Hospital, Lund University, Jan Waldenströms gata 59, SE- 205 02 Malmö, Sweden. *E-mail: [email protected] Accepted May 4, 2013 Epub ahead of print Oct 3, 2013 Serology is a main tool for syphilis diagnostic work used (the Captia Syphilis-Enzywell Treponema IgG and IgM, (1–3). According to a recent survey performed by the respectively, EIA from Diesse, Monteriggioni, Italy); the IgG method uses antigen-coated wells and IgM is determined by U.S. Centers for Disease Control and Prevention (CDC) capture technique; a value over 1.0 is considered to be reactive 56% of infectious disease specialists responded that they (7). Particle agglutination was done at serum dilution 80 with send a rapid plasma reagin (RPR) test to the laboratory no quantitation of the result (TPPA, Fujirebio, Tokyo, Japan). and treat presumptively for syphilis (if result is negative 18% repeat the RPR testing before treating the patient, and 17% treat only if RPR is positive) (4). However, a RESULTS clearly positive reaction may be missing at the first visit Case 1 was tested twice, at 6 days before and 112 of patients with active syphilis (5). Non-treponemal test days after treatment start (Fig. 1, upper panel). The (e.g. RPR and Venereal Disease Research Laboratory treponemal screening value is rising from 4 to a highly (VDRL)) reactivity is known to decrease in response to reactive value of 14, while the non-treponemal VDRL therapy, in contrast to the typically life-long persistence titre decreases from a weak 1 to negativity. -
Medical Bacteriology
LECTURE NOTES Degree and Diploma Programs For Environmental Health Students Medical Bacteriology Abilo Tadesse, Meseret Alem University of Gondar In collaboration with the Ethiopia Public Health Training Initiative, The Carter Center, the Ethiopia Ministry of Health, and the Ethiopia Ministry of Education September 2006 Funded under USAID Cooperative Agreement No. 663-A-00-00-0358-00. Produced in collaboration with the Ethiopia Public Health Training Initiative, The Carter Center, the Ethiopia Ministry of Health, and the Ethiopia Ministry of Education. Important Guidelines for Printing and Photocopying Limited permission is granted free of charge to print or photocopy all pages of this publication for educational, not-for-profit use by health care workers, students or faculty. All copies must retain all author credits and copyright notices included in the original document. Under no circumstances is it permissible to sell or distribute on a commercial basis, or to claim authorship of, copies of material reproduced from this publication. ©2006 by Abilo Tadesse, Meseret Alem All rights reserved. Except as expressly provided above, no part of this publication may be reproduced or transmitted in any form or by any means, electronic or mechanical, including photocopying, recording, or by any information storage and retrieval system, without written permission of the author or authors. This material is intended for educational use only by practicing health care workers or students and faculty in a health care field. PREFACE Text book on Medical Bacteriology for Medical Laboratory Technology students are not available as need, so this lecture note will alleviate the acute shortage of text books and reference materials on medical bacteriology. -
Rapid Syphilis Testing Protocol Wisconsin Department of Health Services STD Control Program
Rapid Syphilis Testing Protocol Wisconsin Department of Health Services STD Control Program P-01832 June 2017 Table of Contents Bureau of Communicable Diseases (BCD) Staff Contact List ........................................................................ 3 Common Acronyms and Terms ..................................................................................................................... 4 Introduction and Background ....................................................................................................................... 5 Syphilis FAQ ................................................................................................................................................... 6 Rapid Syphilis Testing Algorithm ................................................................................................................... 9 Program Requirements ............................................................................................................................... 10 Agency Flow of Services .............................................................................................................................. 15 Rapid Syphilis Testing in Nontraditional or Outreach Settings ................................................................... 15 Syphilis Risk Assessment with Rapid Tests ................................................................................................. 17 Syphilis Health Check Testing Kit ............................................................................................................... -
Blood Culture Sampling Policy for Adult and Paediatric Patients
Blood Culture Sampling Policy for Adult and Version: 7.0 Paediatric Patients Date Issued: 25 July 2018 Review Date: 30 May 2021 Document Type: Policy Contents Page Paragraph Executive Summary / Policy Statement / Flowchart 2 1 Scope and Purpose 2 2 Definitions 2 3 Indications for taking a blood culture and procedure 4 4 Roles and Responsibilities 7 5 Related Trust Policies 8 6 Implementation (including training and dissemination) 8 7 Process for Monitoring Compliance/Effectiveness of 9 this Policy 8 Arrangements for Review of this Policy 9 9 References 9 Appendices Page Appendix A List of equipment needed for peripheral blood culture 10 sampling Appendix B ANTT clinical guideline for Blood Culture Sampling 10 Appendix C ANTT clinical guideline for Blood Culture Sampling 10 from a CVC Device without a Transducer Appendix D ANTT clinical guideline for Blood Culture Sampling 10 from a CVC Device with a Transducer Appendix E Paediatric ANTT guideline for Blood Culture Sampling 10 from a CVC device Document Status This is a controlled document. Whilst this document may be printed, the electronic version posted on the intranet is the controlled copy. Any printed copies of this document are not controlled. As a controlled document, this document should not be saved onto local or network drives but should always be accessed from the intranet. Page 1 of 11 Executive Summary Blood cultures are used to detect the cause of a bloodstream infection. The result provides a guide to the appropriate treatment of the patient. False positives may occur if micro- organisms from a site outside of the bloodstream are introduced into the sample of blood obtained for culture which can then result in inappropriate antibiotic therapy and a waste of healthcare resources.