Preface |
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xix | |
About the Author |
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xxiii | |
Section 1 Philosophy and Approach to Diagnostic Parasitology |
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1 | (18) |
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Neglected Tropical Diseases |
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2 | (1) |
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Why Perform Diagnostic Parasitology Testing? |
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3 | (2) |
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3 | (1) |
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3 | (1) |
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4 | (1) |
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4 | (1) |
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Epidemiologic Considerations |
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4 | (1) |
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Compromised Patients; Potential Sex Bias Regarding Infection Susceptibility; Aging |
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4 | (1) |
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5 | (1) |
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Who Should Perform Diagnostic Parasitology Testing? |
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5 | (1) |
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5 | (1) |
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5 | (1) |
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Where Should Diagnostic Parasitology Testing Be Performed? |
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6 | (1) |
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6 | (1) |
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Outpatient or Referral Setting |
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6 | (1) |
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6 | (1) |
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Physician Office Laboratories |
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6 | (1) |
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Over-the-Counter (Home Care) Testing |
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7 | (1) |
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7 | (1) |
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What Factors Should Precipitate Testing? |
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7 | (1) |
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Travel and Residence History |
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7 | (1) |
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Immune Status of the Patient |
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7 | (1) |
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7 | (1) |
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Documented Previous Infection |
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7 | (1) |
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Contact with Infected Individuals |
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8 | (1) |
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Potential Outbreak Testing |
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8 | (1) |
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8 | (1) |
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8 | (1) |
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What Testing Should Be Performed? |
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8 | (2) |
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8 | (1) |
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Special Testing and Reference Laboratories |
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9 | (1) |
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Specialized Referral Test Options-DPDx and Other Sites |
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9 | (1) |
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Other (Nonmicrobiological) Testing |
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10 | (1) |
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What Factors Should Be Considered in Development of Test Menus? |
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10 | (1) |
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10 | (1) |
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10 | (1) |
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Customer Requirements and Perceived Levels of Service |
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10 | (1) |
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Personnel Availability and Level of Expertise |
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10 | (1) |
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10 | (1) |
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11 | (1) |
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Risk Management Issues Associated with Stat Testing |
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11 | (7) |
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Primary Amebic Meningoencephalitis |
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11 | (3) |
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Granulomatous Amebic Encephalitis and Amebic Keratitis |
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14 | (1) |
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15 | (1) |
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Automated Instrumentation |
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15 | (1) |
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16 | (1) |
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16 | (2) |
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Table 1.1 Common features of the neglected tropical diseases |
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18 | (1) |
Section 2 Parasite Classification and Relevant Body Sites |
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19 | (24) |
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20 | (4) |
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20 | (2) |
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22 | (1) |
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22 | (1) |
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Apicomplexa (Including Coccidia) |
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23 | (1) |
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Microsporidia (Now Classified with the Fungi) |
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23 | (1) |
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Protozoa (Other Body Sites) |
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24 | (2) |
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24 | (1) |
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24 | (1) |
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Apicomplexa (Including Coccidia) |
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24 | (1) |
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Microsporidia (Now Classified with the Fungi) |
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25 | (1) |
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Protozoa (Blood and Tissue) |
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26 | (3) |
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Apicomplexa (Including Sporozoa) |
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26 | (1) |
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26 | (3) |
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29 | (1) |
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29 | (1) |
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Nematodes (Blood and Tissue) |
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30 | (1) |
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31 | (1) |
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32 | (1) |
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33 | (1) |
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Trematodes (Liver and Lungs) |
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33 | (2) |
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35 | (1) |
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35 | (1) |
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36 | (1) |
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Table 2.1 Classification of human parasites |
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37 | (2) |
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Table 2.2 Cosmopolitan distribution of common parasitic infections |
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39 | (1) |
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Table 2.3 Body sites and possible parasites recovered |
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40 | (3) |
Section 3 Collection Options |
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43 | (26) |
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44 | (1) |
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Collection of Fresh Stool Specimens |
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44 | (4) |
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44 | (1) |
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Number of Specimens To Be Collected |
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45 | (2) |
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47 | (1) |
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47 | (1) |
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Specimen Type, Stability, and Need for Preservation |
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47 | (1) |
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Preservation of Stool Specimens |
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48 | (8) |
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Overview of Preservatives |
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48 | (1) |
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49 | (1) |
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Sodium Acetate-Acetic Acid-Formalin (SAF) |
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50 | (1) |
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51 | (1) |
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Schaudinn's Fluid containing PVA (Mercury Base) |
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52 | (1) |
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Schaudinn's Fluid containing PVA (Copper Base, Zinc Base) |
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52 | (1) |
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Single-Vial Collection Systems (Other than SAF) |
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53 | (1) |
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Universal Fixative (Total-Fix) |
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54 | (1) |
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Quality Control for Preservatives |
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54 | (1) |
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Procedure Notes for Use of Preservatives (Stool Fixative Collection Vials) |
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55 | (1) |
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Procedure Limitations for Use of Preservatives (Stool Fixative Collection Vials) |
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55 | (1) |
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56 | (1) |
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Collection and Processing |
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56 | (1) |
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Stat Test Requests and Risk Management Issues |
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57 | (1) |
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Collection of Specimens from Other Body Sites |
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57 | (1) |
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Table 3.1 Fecal specimens for parasites: options for collection and processing |
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58 | (3) |
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Table 3.2 Approaches to stool parasitology: test ordering |
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61 | (1) |
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Table 3.3 Preservatives and procedures commonly used in diagnostic parasitology (stool specimens) |
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62 | (2) |
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Table 3.4 Advantages of thin and thick blood films |
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64 | (1) |
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Table 3.5 Advantages and disadvantages of bully coat films |
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64 | (1) |
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Table 3.6 Potential problems of using EDTA anticoagulant for the preparation of thin and thick blood films |
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65 | (1) |
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Table 3.7 Body sites and possible parasites recovered (trophozoites, cysts, oocysts, spores, adults, larvae, eggs, amastigotes, and trypomastigotes) |
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66 | (3) |
Section 4 Specimen Test Options: Routine Diagnostic Methods and Body Sites |
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69 | (26) |
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Ova and Parasite Examination of Stool Specimens |
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70 | (2) |
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Other Diagnostic Methods for Stool Specimens |
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72 | (2) |
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Culture of Larval-Stage Nematodes |
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72 | (1) |
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Estimation of Worm Burdens through Egg Counts |
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73 | (1) |
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Hatching Test for Schistosome Eggs |
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73 | (1) |
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Screening Stool Samples for Recovery of a Tapeworm Scolex |
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74 | (1) |
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Testing of Other Intestinal Tract Specimens |
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74 | (3) |
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74 | (1) |
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75 | (1) |
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Duodenal Drainage Material |
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76 | (1) |
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Duodenal Capsule Technique (Entero-Test) |
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76 | (1) |
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Urogenital Tract Specimens |
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77 | (1) |
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77 | (1) |
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78 | (1) |
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78 | (1) |
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79 | (2) |
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79 | (1) |
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80 | (1) |
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80 | (1) |
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80 | (1) |
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QBC Microhematocrit Centrifugation Method |
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80 | (1) |
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81 | (1) |
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Membrane Filtration Technique |
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81 | (1) |
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81 | (1) |
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Animal Inoculation and Xenodiagnosis |
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82 | (1) |
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Antibody and Antigen Detection |
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82 | (4) |
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82 | (2) |
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Antigen Detection, Nucleic Acid-Based Tests, and Molecular Panels |
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84 | (1) |
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84 | (1) |
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84 | (2) |
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Table 4.1 Body sites, procedures and specimens, recommended methods and relevant parasites, and comments |
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86 | (8) |
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Table 4.2 Serologic, antigen, and probe tests used in the diagnosis of parasitic infections |
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94 | (1) |
Section 5 Specific Test Procedures and Algorithms |
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95 | (122) |
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96 | (3) |
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Calibration Of The Microscope |
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96 | (3) |
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Ova And Parasite Examination |
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99 | (1) |
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99 | (5) |
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Concentration (Sedimentation And Flotation) |
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104 | (10) |
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Sedimentation Concentration (Formalin-Ethyl Acetate) |
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105 | (4) |
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Sedimentation Concentration Using The Universal Fixative (Total-Fix) |
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109 | (1) |
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Flotation Concentration (Zinc Sulfate) |
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110 | (4) |
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114 | (2) |
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Stains Used In The Permanent Stained Smear |
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116 | (16) |
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Trichrome Stain (Wheatley's Method) |
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116 | (6) |
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Iron Hematoxylin Stain (Spencer-Monroe Method) |
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122 | (5) |
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Iron Hematoxylin Stain (Tompkins-Miller Method) |
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127 | (1) |
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Modified Iron Hematoxylin Stain (Incorporating The Carbol Fuchsin Step) |
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128 | (3) |
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131 | (1) |
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Specialized Stains For Coccidia And Microsporidia |
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132 | (18) |
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Kinyoun's Acid-Fast Stain (Cold Method) |
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132 | (4) |
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Modified Ziehl-Neelsen Acid-Fast Stain (Hot Method) |
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136 | (3) |
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Carbol Fuchsin Negative Stain For Cryptosporidium (W. L. Current) |
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139 | (1) |
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Rapid Safranin Method For Cryptosporidium (D. Baxby) |
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139 | (1) |
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Rapid Safranin Method For Cyclospora, Using A Microwave Oven (Govinda Visvesvara) |
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139 | (1) |
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Auramine O Stain For Aplicomplexa (Including Coccidia) (Thomas Hanscheid) |
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140 | (4) |
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Modified Trichrome Stain For Microsporidia (Weber, Green Counterstain) |
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144 | (2) |
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Modified Trichrome Stain For Microsporidia (Ryan, Blue Counterstain) |
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146 | (3) |
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Modified Trichrome Stain For Microsporidia (Evelyn Kokoskin, Hot Method) |
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149 | (1) |
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Fecal Immunoassays For Intestinal Protozoa |
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150 | (3) |
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150 | (1) |
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151 | (1) |
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151 | (1) |
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151 | (1) |
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Comments On The Performance Of Fecal Immunoassays |
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151 | (1) |
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Enzyme Immunoassays (Antigen Detection, No Centrifugation Recommended) |
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152 | (1) |
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Fluorescence (Visual Identification Of The Organisms, Centrifugation Recommended) |
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152 | (1) |
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Lateral-Flow Cartridges (Antigen Detection, No Centrifugation Recommended) |
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153 | (1) |
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153 | (8) |
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Harada-Mori Filter Paper Strip Culture |
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153 | (2) |
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155 | (3) |
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Agar Plate Culture For Strongyloides Stercoralis |
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158 | (3) |
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Other Methods For Gastrointestinal Tract Specimens |
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161 | (10) |
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Examination For Pinworm (Cellulose Tape Preparations) |
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161 | (2) |
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Sigmoidoscopy Specimens (Direct Wet Smear) |
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163 | (3) |
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Sigmoidoscopy Specimens (Permanent Stained Smear) |
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166 | (2) |
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168 | (3) |
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Methods for Urogenital Tract Specimens |
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171 | (10) |
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171 | (1) |
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171 | (3) |
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174 | (2) |
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Urine Concentration (Centrifugation) |
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176 | (3) |
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Urine Concentration (Nuclepore Membrane Filter) |
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179 | (2) |
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Preparation of Blood Films |
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181 | (9) |
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182 | (2) |
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184 | (2) |
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Combination Thick-Thin Blood Films |
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186 | (1) |
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Risk Management Issues Associated With Blood Films |
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187 | (1) |
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Use Of A Reference Laboratory For Parasite Blood Diagnostic Testing |
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188 | (1) |
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Blood Film Reporting With Additional Report Comments |
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189 | (1) |
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189 | (1) |
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190 | (4) |
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190 | (1) |
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190 | (4) |
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194 | (7) |
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194 | (1) |
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195 | (2) |
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Membrane Filtration Concentration |
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197 | (4) |
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Algorithm 5.1 Procedure for processing fresh stool for the O&P examination |
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201 | (1) |
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Algorithm 5.2 Procedure for processing liquid specimens for the O&P examination |
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202 | (1) |
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Algorithm 5.3 Procedure for processing preserved stool for the O&P examination-two-vial collection kit |
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203 | (1) |
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Algorithm 5.4 Procedure for processing SAF-preserved stool for the O&P examination |
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204 | (1) |
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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) |
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205 | (2) |
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Alternate Method For Smear Preparation Directly From Vial |
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206 | (1) |
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Algorithm 5.6 Use of various fixatives and their recommended stains |
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207 | (1) |
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Algorithm 5.7 Ordering algorithm for laboratory examination for intestinal parasites |
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208 | (1) |
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Algorithm 5.8 Procedure for processing blood specimens for examination |
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209 | (1) |
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Table 5.1 Body sites, specimens, and recommended stains |
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210 | (4) |
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Table 5.2 Approaches to stool parasitology: test ordering |
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214 | (1) |
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Table 5.3 Laboratory test reports: notes and optional comments |
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215 | (1) |
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Table 5.4 Parasitemia determined from conventional light microscopy: clinical correlation |
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216 | (1) |
Section 6 Commonly Asked Questions about Diagnostic Parasitology |
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217 | (48) |
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218 | (11) |
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218 | (3) |
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221 | (1) |
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222 | (5) |
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Stool Immunoassay Options |
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227 | (2) |
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Molecular Test Panels (FDA Cleared) |
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229 | (16) |
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A APTIMA Trichomonas vaginalis Assay |
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230 | (1) |
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B Affirm VPIII Microbial Identification Test |
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231 | (1) |
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C Cepheid Xpert TV Assay for Trichomonas vaginalis from Men and Women |
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232 | (1) |
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D BD MAX Enteric Parasite Panel |
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233 | (1) |
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E BioFire FilmArray Gastrointestinal Panel |
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234 | (2) |
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F Luminex (Verigene II GI Flex Assay; Includes Parasites) |
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236 | (2) |
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G Other Pending Molecular Tests |
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238 | (1) |
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238 | (4) |
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242 | (3) |
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245 | (1) |
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245 | (1) |
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246 | (19) |
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246 | (2) |
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248 | (1) |
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249 | (5) |
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254 | (2) |
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256 | (1) |
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257 | (1) |
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258 | (7) |
Section 7 Parasite Identification |
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265 | (200) |
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266 | (29) |
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266 | (4) |
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Entamoeba histolytica/Entamoeba dispar |
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270 | (3) |
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Comments on Entamoeba moshkovskii and Entamoeba bangladeshi |
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273 | (1) |
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274 | (1) |
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275 | (3) |
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278 | (3) |
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281 | (2) |
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283 | (1) |
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284 | (3) |
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287 | (3) |
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Blastocystis spp. (formerly Blastocystis hominis) |
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290 | (5) |
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Flagellates (Intestinal) Giardia lamblia (G. duodenalis, G. intestinalis) |
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295 | (15) |
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298 | (4) |
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302 | (2) |
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304 | (3) |
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Enteromonas hominis, Retortamonas intestinalis |
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307 | (3) |
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310 | (5) |
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310 | (5) |
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315 | (4) |
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315 | (4) |
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319 | (7) |
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319 | (4) |
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Cystoisospora (formerly Isospora) belli |
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323 | (3) |
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Microsporidia (Intestinal) |
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326 | (8) |
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326 | (3) |
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Encephalitozoon intestinalis, Encephalitozoon spp. |
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329 | (5) |
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Sporozoa (Blood and Tissue) |
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334 | (34) |
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334 | (5) |
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339 | (4) |
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343 | (3) |
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Plasmodium ovale wallickeri, Plasmodium ovale curtisi |
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346 | (4) |
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350 | (3) |
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353 | (2) |
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Babesia spp. (Babesia microti, B. duncani, B. divergens, B. venatorum) |
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355 | (4) |
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359 | (9) |
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Flagellates (Blood and Tissue) |
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368 | (13) |
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368 | (4) |
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Trypanosoma brucei gambiense (West), I brucei rhodesiense (East) |
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372 | (4) |
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376 | (5) |
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Amebae (Other Body Sites) |
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381 | (8) |
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381 | (4) |
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Acanthamoeba spp., Balamuthia mandrillaris, Sappinia diploidea |
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385 | (4) |
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Flagellates (Other Body Sites) |
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389 | (3) |
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389 | (3) |
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392 | (33) |
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392 | (15) |
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392 | (3) |
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Trichuris trichiura, Capillaria philippinensis |
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395 | (3) |
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Necator americanus, Ancylostoma duodenale, Ancylostoma ceylanicum (Hookworms); Trichostrongylus spp. |
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398 | (3) |
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Strongyloides stercoralis |
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401 | (3) |
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404 | (3) |
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407 | (12) |
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Ancylostoma braziliense, Ancylostoma caninum, Uncinaria stenocephala (Dog and Cat Hookworms) |
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407 | (3) |
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Toxocara canis, Toxocara cati (Dog and Cat Ascarid Worms) |
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410 | (3) |
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413 | (3) |
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416 | (3) |
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419 | (6) |
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419 | (6) |
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425 | (23) |
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425 | (18) |
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425 | (3) |
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428 | (4) |
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432 | (3) |
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Hymenolepis (Rodentolepis) nana |
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435 | (3) |
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438 | (2) |
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440 | (3) |
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443 | (5) |
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Echinococcus granulosus, Echinococcus multilocularis, Echinococcus vogeli, Echinococcus oligarthrus |
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443 | (5) |
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448 | (17) |
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448 | (3) |
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448 | (3) |
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451 | (9) |
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Paragonimus westermani, Paragonimus mexicanus, Paragonimus kellicotti |
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451 | (3) |
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454 | (3) |
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Clonorchis (Opisthorchis) sinensis (Chinese liver fluke) |
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457 | (3) |
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460 | (30) |
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Schistosoma spp. (Schistosoma mansoni, S. haematobium, S. japonicum, S. mekongi, S. malayensis, S. intercalatum) |
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460 | (5) |
Section 8 Common Problems in Parasite Identification |
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465 | (24) |
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Table 8.1 Entamoeba sp. trophozoites versus macrophages |
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486 | (1) |
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Table 8.2 Entamoeba sp. cysts versus PMNs |
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486 | (1) |
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Table 8.3 Entamoeba histolytica versus Entamoeba coli precysts and cysts |
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487 | (1) |
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Table 8.4 Endolimax nana versus Dientamoeba fragilis |
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488 | (1) |
Section 9 Identification Aids |
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489 | (58) |
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Diagnostic Considerations |
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490 | (5) |
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Table 9.1 Rapid diagnostic procedures |
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490 | (1) |
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Table 9.2 Diagnostic characteristics for organisms in wet mounts (direct or concentration sediment) |
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491 | (1) |
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Table 9.3 Diagnostic characteristics for organisms in permanent stained smears (e.g., Wheatley's trichrome, iron-hematoxylin) |
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491 | (1) |
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Identification Key 9.1 Identification of intestinal amebae (permanent stained smear) |
|
|
491 | (1) |
|
Identification Key 9.2 Identification of intestinal flagellates |
|
|
492 | (1) |
|
Identification Key 9.3 Identification of helminth eggs |
|
|
493 | (1) |
|
Identification Key 9.4 Identification of microfilariae |
|
|
494 | (1) |
|
|
495 | (17) |
|
Table 9.4 Intestinal protozoa: trophozoites of common amebae |
|
|
495 | (2) |
|
Table 9.5 Intestinal protozoa: cysts of common amebae |
|
|
497 | (2) |
|
Table 9.6 Intestinal protozoa: trophozoites of less common amebae |
|
|
499 | (1) |
|
Table 9.7 Intestinal protozoa: cysts of less common amebae |
|
|
499 | (1) |
|
Table 9.8 Morphologic criteria used to identify Blastocystis spp. |
|
|
500 | (1) |
|
Table 9.9 Intestinal protozoa: trophozoites of flagellates |
|
|
501 | (1) |
|
Table 9.10 Intestinal protozoa: cysts of flagellates |
|
|
502 | (1) |
|
Table 9.11 Intestinal protozoa: ciliate (Balantidium colt) |
|
|
503 | (1) |
|
|
503 | (1) |
|
Table 9.13 Microsporidia (related to the Fungi): general information |
|
|
504 | (1) |
|
Table 9.14 Microsporidia: recommended diagnostic techniques |
|
|
505 | (1) |
|
Table 9.15 Comparison of Naegleria fowleri, Acanthamoeba spp., Balamuthia mandrillaris, and Sappinia dip/oidea |
|
|
506 | (1) |
|
Table 9.16 Characteristics of Trichomonas vaginalis |
|
|
507 | (1) |
|
Table 9.17 Key characteristics of intestinal tract and urogenital system protozoa |
|
|
508 | (4) |
|
|
512 | (20) |
|
Table 9.18 Normal life spans of the most common intestinal nematodes |
|
|
512 | (1) |
|
Table 9.19 Characteristics of the most common intestinal nematodes |
|
|
513 | (2) |
|
Table 9.20 Tissue nematodes |
|
|
515 | (2) |
|
Table 9.21 Trichinella spiralis: life cycle stages and clinical conditions |
|
|
517 | (1) |
|
Table 9.22 Characteristics of human microfilariae |
|
|
518 | (1) |
|
Table 9.23 Characteristics of cestode parasites (intestinal) |
|
|
519 | (2) |
|
Table 9.24 Tissue cestodes |
|
|
521 | (2) |
|
Table 9.25 Characteristics of intestinal trematodes |
|
|
523 | (1) |
|
Table 9.26 Characteristics of liver and lung trematodes |
|
|
524 | (2) |
|
Table 9.27 Human paragonimiasis |
|
|
526 | (1) |
|
Table 9.28 Characteristics of blood trematodes |
|
|
527 | (1) |
|
Table 9.29 Key characteristics of helminths |
|
|
528 | (4) |
|
|
532 | (15) |
|
Table 9.30 Malaria characteristics with fresh blood or EDTA-blood |
|
|
532 | (1) |
|
Table 9.31 Potential problems with using EDTA anticoagulant for the preparation of thin and thick blood films |
|
|
533 | (1) |
|
Table 9.32 Plasmodia in Giemsa-stained thin blood smears |
|
|
534 | (3) |
|
Table 9.33 Relevant issues for handling requests for identification of infectious blood parasites |
|
|
537 | (3) |
|
Table 9.34 Features of human leishmanial infections |
|
|
540 | (1) |
|
Table 9.35 Characteristics of American trypanosomiasis |
|
|
541 | (1) |
|
Table 9.36 Characteristics of East and West African trypanosomiasis |
|
|
541 | (1) |
|
Table 9.37 Key characteristics of blood parasites |
|
|
541 | (6) |
Index |
|
547 | |