
Practical Guide to Diagnostic Parasitology
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Specimen collection, preservation, and testing options are thoroughly discussed, from the routine ova and parasite examination to blood films, fecal immunoassays, and the newer molecular test panels. Specific test procedures, laboratory methods and reagents, and algorithms are provided. The ever-helpful 'FAQ' section of commonly asked questions now offers expanded information on stool specimen fixatives and testing, thorough coverage of new techniques, and advice on reporting and commenting on results.
The heart of the Guide, covering identification of individual pathogens, has been expanded with more discussion and comparison of organisms and dozens of new color images. An entirely new section has been added that uses extensive figures and new tables to illustrate common problems with differentiating organisms from one another and from possible microscopic artifacts. The final section has been reorganized to include identification keys and dozens of tables summarizing organism characteristics to assist the bench microbiologist with routine diagnostic testing methods.
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Content
- Cover
- Title Page
- Copyright Page
- Dedication Page
- Contents
- Preface
- About the Author
- Section 1 Philosophy and Approach to Diagnostic Parasitology
- Neglected Tropical Diseases
- Why Perform Diagnostic Parasitology Testing?
- Travel
- Population Movements
- Control Issues
- Climate Change
- Epidemiologic Considerations
- Compromised Patients
- Potential Sex Bias Regarding Infection Susceptibility
- Aging
- Approach to Therapy
- Who Should Perform Diagnostic Parasitology Testing?
- Laboratory Personnel
- Nonlaboratory Personnel
- Where Should Diagnostic Parasitology Testing Be Performed?
- Inpatient Setting
- Outpatient or Referral Setting
- Decentralized Testing
- Physician Office Laboratories
- Over-the-Counter (Home Care) Testing
- Field Sites
- What Factors Should Precipitate Testing?
- Travel and Residence History
- Immune Status of the Patient
- Clinical Symptoms
- Documented Previous Infection
- Contact with Infected Individuals
- Potential Outbreak Testing
- Occupational Testing
- Therapeutic Failure
- What Testing Should Be Performed?
- Routine Tests
- Special Testing and Reference Laboratories
- Specialized Referral Test Options-DPDx and Other Sites
- Other (Nonmicrobiological) Testing
- What Factors Should Be Considered in Development of Test Menus?
- Physical Plant
- Client Base
- Customer Requirements and Perceived Levels of Service
- Personnel Availability and Level of Expertise
- Equipment
- Budget
- Risk Management Issues Associated with Stat Testing
- Primary Amebic Meningoencephalitis
- Granulomatous Amebic Encephalitis and Amebic Keratitis
- Request for Blood Films
- Automated Instrumentation
- Patient Information
- Conventional Microscopy
- References
- Table 1.1 Common features of the neglected tropical diseases
- Section 2 Parasite Classificationand Relevant Body Sites
- Protozoa (Intestinal)
- Amebae, Stramenopiles
- Flagellates
- Ciliates
- Apicomplexa (Including Coccidia)
- Microsporidia (Now Classified with the Fungi)
- Protozoa (Other Body Sites)
- Amebae
- Flagellates
- Apicomplexa (Including Coccidia)
- Microsporidia (Now Classified with the Fungi)
- Protozoa (Blood and Tissue)
- Apicomplexa (Including Sporozoa)
- Flagellates
- Nematodes (Intestinal)
- Nematodes (Tissue)
- Nematodes (Blood and Tissue)
- Cestodes (Intestinal)
- Cestodes (Tissue)
- Trematodes (Intestinal)
- Trematodes (Liver and Lungs)
- Trematodes (Blood)
- Pentastomids
- Acanthocephala
- Suggested Reading
- Table 2.1 Classification of human parasites
- Table 2.2 Cosmopolitan distribution of common parasitic infections
- Table 2.3 Body sites and possible parasites recovered
- Section 3 Collection Options
- Safety
- Collection of Fresh Stool Specimens
- Collection Method
- Number of Specimens To Be Collected
- Collection Times
- Posttherapy Collection
- Specimen Type, Stability, and Need for Preservation
- Preservation of Stool Specimens
- Overview of Preservatives
- Formalin
- Sodium Acetate-Acetic Acid-Formalin (SAF)
- Schaudinn's Fluid
- Schaudinn's Fluid Containing PVA (Mercury Base)
- Schaudinn's Fluid Containing PVA (Copper Base, Zinc Base)
- Single-Vial Collection Systems (Other than SAF)
- Universal Fixative (Total-Fix)
- Quality Control for Preservatives
- Procedure Notes for Use of Preservatives (Stool Fixative Collection Vials)
- Procedure Limitations for Use of Preservatives (Stool Fixative Collection Vials)
- Collection of Blood
- Collection and Processing
- Stat Test Requests and Risk Management Issues
- Collection of Specimens from Other Body Sites
- Suggested Reading
- Table 3.1 Fecal specimens for parasites: options for collection and processing
- Table 3.2 Approaches to stool parasitology: test ordering
- Table 3.3 Preservatives and procedures commonly used in diagnostic parasitology (stool specimens)
- Table 3.4 Advantages of thin and thick blood films
- Table 3.5 Advantages and disadvantages of buffy coat films
- Table 3.6 Potential problems of using EDTA anticoagulant for the preparation of thin and thick blood films
- Table 3.7 Body sites and possible parasites recovered (trophozoites, cysts, oocysts, spores, adults, larvae, eggs, amastigotes, and trypomastigotes)
- Section 4 Specimen Test Options: Routine Diagnostic Methods and Body Sites
- Ova and Parasite Examination of Stool Specimens
- Other Diagnostic Methods for Stool Specimens
- Culture of Larval-Stage Nematodes
- Estimation of Worm Burdens through Egg Counts
- Hatching Test for Schistosome Eggs
- Screening Stool Samples for Recovery of a Tapeworm Scolex
- Testing of Other Intestinal Tract Specimens
- Examination for Pinworm
- Sigmoidoscopy Material
- Duodenal Drainage Material
- Duodenal Capsule Technique (Entero-Test)
- Urogenital Tract Specimens
- Sputum
- Aspirates
- Biopsy Specimens
- Blood
- Thin Blood Films
- Thick Blood Films
- Blood Staining Methods
- Buffy Coat Films
- QBC Microhematocrit Centrifugation Method
- Knott Concentration
- Membrane Filtration Technique
- Culture Methods
- Animal Inoculation and Xenodiagnosis
- Antibody and Antigen Detection
- Antibody Detection
- Antigen Detection, Nucleic Acid-Based Tests, and Molecular Panels
- Intradermal Tests
- UV Autofluorescence
- Suggested Reading
- Table 4.1 Body sites, procedures and specimens, recommended methods and relevant parasites, and comments
- Table 4.2 Serologic, antigen, and probe tests used in the diagnosis of parasitic infections
- Section 5 Specific Test Procedures and Algorithms
- Microscopy
- CALIBRATION OF THE MICROSCOPE
- Ova and Parasite Examination
- DIRECT WET FECAL SMEAR
- CONCENTRATION (Sedimentation and Flotation)
- SEDIMENTATION CONCENTRATION (Formalin-Ethyl Acetate)
- SEDIMENTATION CONCENTRATION USING THE UNIVERSAL FIXATIVE (Total-Fix)
- FLOTATION CONCENTRATION (Zinc Sulfate)
- PERMANENT STAINED SMEAR
- Stains Used in the Permanent Stained Smear
- TRICHROME STAIN (Wheatley's Method)
- IRON HEMATOXYLIN STAIN (Spencer-Monroe Method)
- IRON HEMATOXYLIN STAIN (Tompkins-Miller Method)
- MODIFIED IRON HEMATOXYLIN STAIN (Incorporating the Carbol Fuchsin Step)
- CHLORAZOL BLACK E STAIN
- Specialized Stains for Coccidia and Microsporidia
- KINYOUN'S ACID-FAST STAIN (Cold Method)
- MODIFIED ZIEHL-NEELSEN ACID-FAST STAIN (Hot Method)
- CARBOL FUCHSIN NEGATIVE STAIN FOR CRYPTOSPORIDIUM (W. L. Current)
- RAPID SAFRANIN METHOD FOR CRYPTOSPORIDIUM (D. Baxby)
- RAPID SAFRANIN METHOD FOR CYCLOSPORA, USING A MICROWAVE OVEN (Govinda Visvesvara)
- AURAMINE O STAIN FOR APICOMPLEXA (INCLUDING COCCIDIA) (Thomas Hänscheid)
- MODIFIED TRICHROME STAIN FOR MICROSPORIDIA (Weber, Green Counterstain)
- MODIFIED TRICHROME STAIN FOR MICROSPORIDIA (Ryan, Blue Counterstain)
- MODIFIED TRICHROME STAIN FOR MICROSPORIDIA (Evelyn Kokoskin, Hot Method)
- Fecal Immunoassays for Intestinal Protozoa
- ENTAMOEBA HISTOLYTICA
- CRYPTOSPORIDIUM SPP.
- GIARDIA LAMBLIA
- KITS UNDER DEVELOPMENT
- COMMENTS ON THE PERFORMANCE OF FECAL IMMUNOASSAYS
- ENZYME IMMUNOASSAYS (Antigen Detection, No Centrifugation Recommended)
- FLUORESCENCE (Visual Identification of the Organisms, Centrifugation Recommended)
- LATERAL-FLOW CARTRIDGES (Antigen Detection, No Centrifugation Recommended)
- Larval Nematode Culture
- HARADA-MORI FILTER PAPER STRIP CULTURE
- BAERMANN CONCENTRATION
- AGAR PLATE CULTURE FOR STRONGYLOIDES STERCORALIS
- Other Methods for Gastrointestinal Tract Specimens
- EXAMINATION FOR PINWORM (Cellulose Tape Preparations)
- SIGMOIDOSCOPY SPECIMENS (Direct Wet Smear)
- SIGMOIDOSCOPY SPECIMENS (Permanent Stained Smear)
- DUODENAL ASPIRATES
- Methods for Urogenital Tract Specimens
- RECEIPT OF DRY SMEARS
- DIRECT SALINE MOUNT
- PERMANENT STAINED SMEAR
- URINE CONCENTRATION (Centrifugation)
- URINE CONCENTRATION (Nuclepore Membrane Filter)
- Preparation of Blood Films
- THIN BLOOD FILMS
- THICK BLOOD FILMS
- COMBINATION THICK-THIN BLOOD FILMS
- RISK MANAGEMENT ISSUES ASSOCIATED WITH BLOOD FILMS
- USE OF A REFERENCE LABORATORY FOR PARASITE BLOOD DIAGNOSTIC TESTING
- BLOOD FILM REPORTING WITH ADDITIONAL REPORT COMMENTS
- BUFFY COAT BLOOD FILMS
- Blood Stains
- STAIN OPTIONS
- GIEMSA STAIN
- Blood Concentration
- BUFFY COAT CONCENTRATION
- KNOTT CONCENTRATION
- MEMBRANE FILTRATION CONCENTRATION
- Suggested Reading
- Algorithm 5.1 Procedure for processing fresh stool for the O&P examination
- Algorithm 5.2 Procedure for processing liquid specimens for the O&P examination
- Algorithm 5.3 Procedure for processing preserved stool for the O&P examination-two-vial collection kit
- Algorithm 5.4 Procedure for processing SAF-preserved stool for the O&P examination
- Algorithm 5.5 Procedure for the use of Total-Fix (universal fixative, single-vial system) (this fixative contains no mercury, no PVA, and no formalin)
- Algorithm 5.6 Use of various fixatives and their recommended stains
- Algorithm 5.7 Ordering algorithm for laboratory examination for intestinal parasites
- Algorithm 5.8 Procedure for processing blood specimens for examination
- Table 5.1 Body sites, specimens, and recommended stains
- Table 5.2 Approaches to stool parasitology: test ordering
- Table 5.3 Laboratory test reports: notes and optional comments
- Table 5.4 Parasitemia determined from conventional light microscopy: clinical correlation
- Section 6 Commonly Asked Questions about Diagnostic Parasitology
- Stool Parasitology
- Specimen Collection
- Specimen Processing
- Diagnostic Methods
- Stool Immunoassay Options
- MOLECULAR TEST PANELS (FDA CLEARED)
- A. APTIMA Trichomonas vaginalis Assay
- B. Affirm VPIII Microbial Identification Test
- C. Cepheid Xpert TV Assay for Trichomonas vaginalis from Men and Women
- D. BD MAX Enteric Parasite Panel
- E. BioFire FilmArray Gastrointestinal Panel
- F. Luminex (Verigene II GI Flex Assay
- Includes Parasites)
- G. Other Pending Molecular Tests
- Organism Identification
- Reporting
- Proficiency Testing
- Tissues or Fluids
- Blood
- Specimen Collection
- Specimen Processing
- Diagnostic Methods
- Organism Identification
- Reporting
- Proficiency Testing
- General Questions
- Suggested Reading
- Section 7 Parasite Identification
- PROTOZOA
- Amebae (Intestinal)
- Entamoeba histolytica
- Entamoeba histolytica/Entamoeba dispar
- Comments on Entamoeba moshkovskii and Entamoeba bangladeshi
- Entamoeba bangladeshi
- Entamoeba hartmanni
- Entamoeba coli
- Entamoeba gingivalis
- Entamoeba polecki
- Endolimax nana
- Iodamoeba bütschlii
- Blastocystis spp. (formerly Blastocystis hominis)
- Flagellates (Intestinal)
- Giardia lamblia (G. duodenalis, G. intestinalis)
- Dientamoeba fragilis
- Chilomastix mesnili
- Pentatrichomonas hominis
- Enteromonas hominis, Retortamonas intestinalis
- Ciliates (Intestinal)
- Balantidium coli
- Apicomplexa (Intestinal)
- Cryptosporidium spp.
- Coccidia (Intestinal)
- Cyclospora cayetanensis
- Cystoisospora (formerly Isospora) belli
- Microsporidia (Intestinal)
- Enterocytozoon bieneusi
- Encephalitozoon intestinalis, Encephalitozoon spp.
- Sporozoa (Blood and Tissue)
- Plasmodium vivax
- Plasmodium falciparum
- Plasmodium malariae
- Plasmodium ovale wallickeri, Plasmodium ovale curtisi
- Plasmodium knowlesi
- Malaria
- Babesia spp. (Babesia microti, B. duncani, B. divergens, B. venatorum)
- Toxoplasma gondii
- Flagellates (Blood and Tissue)
- Leishmania spp.
- Trypanosoma brucei gambiense (West), T. brucei rhodesiense (East)
- Trypanosoma cruzi
- Amebae (Other Body Sites)
- Naegleria fowleri
- Acanthamoeba spp., Balamuthia mandrillaris, Sappinia diploidea
- Flagellates (Other Body Sites)
- Trichomonas vaginalis
- NEMATODES
- Intestinal
- Ascaris lumbricoides
- Trichuris trichiura, Capillaria philippinensis
- Necator americanus, Ancylostoma duodenale, Ancylostoma ceylanicum (Hookworms)
- Trichostrongylus spp.
- Strongyloides stercoralis
- Enterobius vermicularis
- Tissue
- Ancylostoma braziliense, Ancylostoma caninum, Uncinaria stenocephala, (Dog and Cat Hookworms)
- Toxocara canis, Toxocara cati, (Dog and Cat Ascarid Worms)
- Dracunculus medinensis
- Trichinella spiralis
- Blood and Tissue
- Filarial Worms
- CESTODES
- Intestinal
- Taenia saginata
- Taenia solium
- Diphyllobothrium latum
- Hymenolepis (Rodentolepis) nana
- Hymenolepis diminuta
- Dipylidium caninum
- Tissue
- Echinococcus granulosus, Echinococcus multilocularis, Echinococcus vogeli, Echinococcus oligarthrus
- TREMATODES
- Intestinal
- Fasciolopsis buski
- Liver and Lungs
- Paragonimus westermani, Paragonimus mexicanus, Paragonimus kellicotti
- Fasciola hepatica
- Clonorchis (Opisthorchis) sinensis (Chinese liver fluke)
- Blood
- Schistosoma spp. (Schistosoma mansoni, S. haematobium, S. japonicum, S. mekongi, S. malayensis, S. intercalatum)
- Suggested Reading
- Section 8 Common Problems in Parasite Identification
- Table 8.1 Entamoeba sp. trophozoites versus macrophages
- Table 8.2 Entamoeba sp. cysts versus PMNs
- Table 8.3 Entamoeba histolytica versus Entamoeba coli precysts and cysts
- Table 8.4 Endolimax nana versus Dientamoeba fragilis
- Section 9 Identification Aids
- DIAGNOSTIC CONSIDERATIONS
- Table 9.1 Rapid diagnostic procedures
- Table 9.2 Diagnostic characteristics for organisms in wet mounts (direct or concentration sediment)
- Table 9.3 Diagnostic characteristics for organisms in permanent stained smears (e.g., Wheatley's trichrome, iron-hematoxylin)
- Identification Key 9.1 Identification of intestinal amebae (permanent stained smear)
- Identification Key 9.2 Identification of intestinal flagellates
- Identification Key 9.3 Identification of helminth eggs
- Identification Key 9.4 Identification of microfilariae
- PROTOZOA
- Table 9.4 Intestinal protozoa: trophozoites of common amebae
- Table 9.5 Intestinal protozoa: cysts of common amebae
- Table 9.6 Intestinal protozoa: trophozoites of less common amebae
- Table 9.7 Intestinal protozoa: cysts of less common amebae
- Table 9.8 Morphologic criteria used to identify Blastocystis spp.
- Table 9.9 Intestinal protozoa: trophozoites of flagellates
- Table 9.10 Intestinal protozoa: cysts of flagellates
- Table 9.11 Intestinal protozoa: ciliate (Balantidium coli)
- Table 9.12 Apicomplexa
- Table 9.13 Microsporidia (related to the Fungi): general information
- Table 9.14 Microsporidia: recommended diagnostic techniques
- Table 9.15 Comparison of Naegleria fowleri, Acanthamoeba spp., Balamuthia mandrillaris, and Sappinia diploidea
- Table 9.16 Characteristics of Trichomonas vaginalis
- Table 9.17 Key characteristics of intestinal tract and urogenital system protozoa
- HELMINTHS
- Table 9.18 Normal life spans of the most common intestinal nematodes
- Table 9.19 Characteristics of the most common intestinal nematodes
- Table 9.20 Tissue nematodes
- Table 9.21 Trichinella spiralis: life cycle stages and clinical conditions
- Table 9.22 Characteristics of human microfilariae
- Table 9.23 Characteristics of cestode parasites (intestinal)
- Table 9.24 Tissue cestodes
- Table 9.25 Characteristics of intestinal trematodes
- Table 9.26 Characteristics of liver and lung trematodes
- Table 9.27 Human paragonimiasis
- Table 9.28 Characteristics of blood trematodes
- Table 9.29 Key characteristics of helminths
- BLOOD PARASITES
- Table 9.30 Malaria characteristics with fresh blood or EDTA-blood
- Table 9.31 Potential problems with using EDTA anticoagulant for the preparation of thin and thick blood films
- Table 9.32 Plasmodia in Giemsa-stained thin blood smears
- Table 9.33 Relevant issues for handling requests for identification of infectious blood parasites
- Table 9.34 Features of human leishmanial infections
- Table 9.35 Characteristics of American trypanosomiasis
- Table 9.36 Characteristics of East and West African trypanosomiasis
- Table 9.37 Key characteristics of blood parasites
- Index
- EULA
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