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Lynne Shore Garcia. Practical Guide to Diagnostic Parasitology
Contents
Preface
Acknowledgments
SECTION 1 Philosophy and Approach to Diagnostic Parasitology. Why Perform This Type of Testing? Travel
Population Movements
Control Issues
Global Warming
Epidemiologic Considerations
Compromised Patients
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
Other (Nonmicrobiological) Testing
What Factors Should Be Considered When Developing 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
SECTION 2 Parasite Classification and Relevant Body Sites
Protozoa (Intestinal) Amebae
Flagellates
Ciliates
Coccidia
Microsporidia
Protozoa (Other Body Sites) Amebae
Flagellates
Coccidia
Microsporidia
Protozoa (Blood and Tissue) Sporozoa
Flagellates. Leishmaniae
Trypanosomes
Nematodes (Intestinal)
Nematodes (Tissue)
Nematodes (Blood and Tissue)
Cestodes (Intestinal)
Cestodes (Tissue)
Trematodes (Intestinal)
Trematodes (Liver and Lungs)
Trematodes (Blood)
Pentastomids
Acanthocephala
Suggested Reading
SECTION 3. Collection Options
Safety
Collection of Fresh Stool Specimens
Collection Method
Number of Specimens To Be Collected. Standard Approach
Different Approaches
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
Polyvinyl Alcohol (PVA)
Modified PVA (Mercury Substitutes)
Single-Vial Collection Systems (Other Than SAF)
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
Suggested Reading
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 and Nucleic Acid-Based Tests
Intradermal Tests
Suggested Reading
SECTION 5. Specific Test Procedures and Algorithms. Microscopy. CALIBRATION OF THE MICROSCOPE. Description
Supplies
Equipment
Quality Control
Detailed Procedure
Reporting
Procedure Reminders
Procedure Limitations
Ova and Parasite Examination. DIRECT WET FECAL SMEAR. Description
Reagents
Quality Control
Detailed Procedure
Reporting
Procedure Reminders
Procedure Limitations
Concentration (Sedimentation and Flotation) Description
Commercial Devices
SEDIMENTATION CONCENTRATION (Formalin-Ethyl Acetate)
Reagents
Quality Control
Detailed Procedure
Reporting
Procedure Reminders
Procedure Limitations
FLOTATION CONCENTRATION (Zinc Sulfate) Description
Reagents
Quality Control
Detailed Procedure
Reporting
Procedure Reminders
Procedure Limitations
PERMANENT STAINED SMEAR. Description
Preparation of Material for Staining
Stains Used in the Permanent Stained Smear. TRICHROME STAIN (Wheatley’s Method) Description
Reagents
Quality Control
Detailed Procedure for Using Mercury-Based Fixatives
Detailed Procedure for Using Non-Mercury-Based Fixatives (Iodine-Alcohol Step and Alcohol Rinse Not Required)
Reporting
Procedure Reminders
Procedure Limitations
IRON HEMATOXYLIN STAIN (Spencer-Monroe Method) Description
Reagents
Quality Control
Detailed Procedure for Using Mercury-Based Fixatives
Detailed Procedure for Using Non-Mercury-Based Fixatives
Reporting
Procedure Reminders
Procedure Limitations
IRON HEMATOXYLIN STAIN (Tompkins-Miller Method) Description
Detailed Procedure
MODIFIED IRON HEMATOXYLIN STAIN (Incorporating the Carbol Fuchsin Step) Description
Reagents
Detailed Procedure
Procedure Reminders
POLYCHROME IV STAIN
CHLORAZOL BLACK E STAIN
Specialized Stains for Coccidia and Microsporidia. KINYOUN’S ACID-FAST STAIN (Cold Method) Description
Reagents
Quality Control
Detailed Procedure
Reporting
Procedure Reminders
Procedure Limitations
MODIFIED ZIEHL-NEELSEN ACID-FAST STAIN (Hot Method) Description
Reagents
Quality Control
Detailed Procedure
Reporting
Procedure Reminders
Procedure Limitations
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 COCCIDIA (Thomas Hänscheid)
Quality Control
Procedure
Reporting
Procedure Notes
Procedure Limitations
MODIFIED TRICHROME STAIN FOR MICROSPORIDIA (Weber, Green Counterstain) Description
Reagents
Quality Control
Detailed Procedure
Reporting
Procedure Reminders
Procedure Limitations
MODIFIED TRICHROME STAIN FOR MICROSPORIDIA (Ryan, Blue Counterstain) Description
Reagents
Quality Control
Detailed Procedure
Reporting
Procedure Reminders
Procedure Limitations
MODIFIED TRICHROME STAIN FOR MICROSPORIDIA (Evelyn Kokoskin, Hot Method) Description
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)
Lateral-Flow Cartridges (Antigen Detection, No Centrifugation Recommended)
Larval Nematode Culture. HARADA-MORI FILTER PAPER STRIP CULTURE. Description
Quality Control
Detailed Procedure
Reporting
Procedure Reminders
Procedure Limitations
BAERMANN CONCENTRATION. Description
Quality Control
Detailed Procedure
Reporting
Procedure Reminders
Procedure Limitations
AGAR PLATE CULTURE FOR STRONGYLOIDES STERCORALIS. Description
Reagents
Quality Control
Reporting
Procedure Reminders
Procedure Limitations
Other Methods for Gastrointestinal Tract Specimens. EXAMINATION FOR PINWORM (Cellulose Tape Preparations) Description
Quality Control
Detailed Procedure
Reporting
Procedure Reminders
Procedure Limitations
SIGMOIDOSCOPY SPECIMENS (Direct Wet Smear) Description
Reagents
Detailed Procedure
Reporting
Procedure Reminders
Procedure Limitations
SIGMOIDOSCOPY SPECIMENS (Permanent Stained Smear) Description
Reagents
Quality Control
Detailed Procedure
Reporting
Procedure Reminders
Procedure Limitations
DUODENAL ASPIRATES. Description
Reagents
Quality Control
Detailed Procedure
Reporting
Procedure Reminders
Procedure Limitations
Methods for Urogenital Tract Specimens. RECEIPT OF DRY SMEARS
DIRECT SALINE MOUNT. Description
Reagents
Quality Control
Detailed Procedure
Reporting
Procedure Reminders
Procedure Limitations
PERMANENT STAINED SMEAR. Description
Reagents
Quality Control
Detailed Procedure
Reporting
Procedure Reminders
Procedure Limitations
URINE CONCENTRATION (Centrifugation) Description
Reagents
Quality Control
Detailed Procedure
Reporting
Procedure Reminders
Procedure Limitations
URINE CONCENTRATION (Nuclepore Membrane Filter) Description
Reagents
Quality Control
Detailed Procedure
Reporting
Procedure Reminders
Procedure Limitations
Preparation of Blood Films
THIN BLOOD FILMS. Description
Reagents
Quality Control
Detailed Procedure
Procedure Reminders
Procedure Limitations
THICK BLOOD FILMS. Description
Quality Control
Detailed Procedure
Procedure Reminders
Procedure Limitations
COMBINATION THICK-THIN BLOOD FILMS. Description
Quality Control
Detailed Procedure
Procedure Reminders
Procedure Limitations
BUFFY COAT BLOOD FILMS
Blood Stains
GIEMSA STAIN. Description
Reagents
Quality Control
Detailed Procedure
Thick Blood Films (Only)
Reporting
Procedure Reminders
Procedure Limitations
Blood Concentration
BUFFY COAT CONCENTRATION. Description
Reagents
Quality Control
Detailed Procedure
Reporting
Procedure Reminders
Procedure Limitations
KNOTT CONCENTRATION. Description
Reagents
Quality Control
Detailed Procedure
Reporting
Procedure Reminders
Procedure Limitations
MEMBRANE FILTRATION CONCENTRATION. Description
Reagents
Quality Control
Detailed Procedure
Reporting
Procedure Reminders
Procedure Limitations
Suggested Reading
SECTION 6. Commonly Asked Questions about Diagnostic Parasitology
Stool Parasitology. Specimen Collection. Intestinal Tract
Fixatives
Specimen Processing. O&P Exam
Diagnostic Methods. Direct Wet Examinations
Concentrations
Permanent Stains
Stool Immunoassay Options
Organism Identification. Protozoa
Helminths
Reporting. Organism Identification
Quantitation
Proficiency Testing. Wet Preparations
Permanent Stained Smears
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
Diagnosis
General Comments
Description (Trophozoite)
Description (Cyst)
Entamoeba dispar
Diagnosis
General Comments
Description (Trophozoite)
Description (Cyst)
Entamoeba hartmanni
Diagnosis
General Comments
Description (Trophozoite)
Description (Cyst)
Entamoeba coli
Diagnosis
General Comments
Description (Trophozoite)
Description (Cyst)
Entamoeba gingivalis
Comments
Entamoeba polecki
Comments
Description
Description
Endolimax nana
Diagnosis
General Comments
Description (Trophozoite)
Description (Cyst)
Iodamoeba bütschlii
Diagnosis
General Comments
Description (Trophozoite)
Description (Cyst)
Blastocystis hominis
Diagnosis
General Comments
Description
Additional Information
Flagellates (Intestinal) Giardia lamblia
Diagnosis
General Comments
Description (Trophozoite)
Description (Cyst)
Dientamoeba fragilis
Diagnosis
General Comments
Description
Additional Information
Chilomastix mesnili
Diagnosis
General Comments
Description (Trophozoite)
Description (Cyst)
Pentatrichomonas hominis
Diagnosis
General Comments
Description
Additional Information
Trichomonas tenax
Enteromonas hominis Retortamonas intestinalis
Diagnosis
General Comments
Description (Trophozoite)
Description (Cyst)
Ciliates (Intestinal) Balantidium coli
Diagnosis
General Comments
Description (Trophozoite)
Description (Cyst)
Additional Information
Coccidia (Intestinal) Cryptosporidium spp
Diagnosis
General Comments
Description
Additional Information
Cyclospora cayetanensis
Diagnosis
General Comments
Additional Information
Isospora (Cystoisospora) belli
Diagnosis
General Comments
Additional Information
Microsporidia (Intestinal) Enterocytozoon bieneusi
Diagnosis
General Comments
Additional Information
Encephalitozoon intestinalis Encephalitozoon spp
Diagnosis
General Comments
Description
Additional Information
Sporozoa (Blood and Tissue) Plasmodium vivax
Diagnosis
General Comments
Additional Information
Plasmodium falciparum
Diagnosis
General Comments
Additional Information
Plasmodium malariae
Diagnosis
General Comments
Additional Information
Plasmodium ovale
Diagnosis
General Comments
Additional Information
Babesia spp
Diagnosis
General Comments
Additional Information
Toxoplasma gondii
Diagnosis
General Comments
Additional Information
Flagellates (Blood and Tissue) Leishmania spp
Diagnosis
General Comments
Additional Information
Trypanosoma brucei gambiense (West) T. brucei rhodesiense (East)
Diagnosis
General Comments
Additional Information
Trypanosoma cruzi
Diagnosis
Life Cycle
Additional Information
Amebae (Other Body Sites) Naegleria fowleri
Diagnosis
General Comments
Description
Additional Information
Amebae (Other Body Sites) Acanthamoeba spp. Balamuthia mandrillaris Sappinia diploidea
Diagnosis
General Comments
Description (Trophozoite)
Description (Cyst)
Flagellates (Other Body Sites) Trichomonas vaginalis
Diagnosis
General Comments
Additional Information
NEMATODES. Intestinal. Ascaris lumbricoides
Diagnosis
General Comments
Description
Additional Information
Intestinal. Trichuris trichiura
Diagnosis
General Comments
Description (Eggs)
Description (Adults)
Additional Information
Necator americanus Ancylostoma duodenale (Hookworms)
Diagnosis
General Comments
Description (Eggs)
Description (Adults)
Additional Information
Strongyloides stercoralis
Diagnosis
General Comments
Description
Additional Information
Enterobius vermicularis
Diagnosis
General Comments
Description (Eggs)
Description (Adults)
Additional Information
Tissue. Ancylostoma braziliense Ancylostoma caninum (Dog and Cat Hookworms)
Diagnosis
General Comments
Additional Information
Toxocara canis Toxocara cati (Dog and Cat Ascarid Worms)
Diagnosis
Life Cycle
Additional Information
Trichinella spiralis
Diagnosis
General Comments
Additional Information
Blood and Tissue. Filarial Worms
Diagnosis
General Comments
Additional Information
CESTODES. Intestinal. Taenia saginata
Diagnosis
General Comments
Description (Eggs)
Description (Adults)
Additional Information
Intestinal. Taenia solium
Diagnosis
General Comments
Description (Eggs and Larvae)
Description (Adult)
Adult-Onset Epilepsy
Diphyllobothrium latum
Diagnosis
General Comments
Description (Eggs)
Additional Information
Description (Adults)
Hymenolepis nana
Diagnosis
General Comments
Description (Eggs)
Description (Adults)
Additional Information
Hymenolepis diminuta
Diagnosis
General Comments
Description (Eggs)
Additional Information
Description (Adults)
Dipylidium caninum
Diagnosis
General Comments
Description (Eggs)
Additional Information
Description (Adults)
Tissue. Echinococcus granulosus
Diagnosis
General Comments
Additional Information
TREMATODES. Intestinal. Fasciolopsis buski
Diagnosis
General Comments
Description
Additional Information
Liver and Lungs. Paragonimus westermani. Paragonimus mexicanus. Paragonimus kellicotti
Diagnosis
General Comments
Description
Additional Information
Fasciola hepatica
Diagnosis
General Comments
Description
Additional Information
Clonorchis sinensis (Opisthorchis sinensis)
Diagnosis
General Comments
Description
Additional Information
Aquaculture
Blood. Schistosoma spp. (S. mansoni, S. haematobium, S. japonicum, S. mekongi, S. intercalatum)
Diagnosis
General Comments
Additional Information
SECTION 8. Identification Aids
Index
