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Chapter 135 : General Approaches for Detection and Identification of Parasites

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Abstract:

This chapter covers various approaches and diagnostic methods currently used for the diagnosis of parasitic infections. If clinical specimens have been properly collected and processed according to specific specimen rejection and acceptance criteria, the examination of prepared wet mounts, concentrated specimens, permanent stained smears, blood films, and various culture materials provides detailed information leading to parasite identification and confirmation of the suspected etiologic agent. Although other tests, such as immunoassay and amplification-based diagnostic kits, continue to become available commercially, the majority of medical parasitology diagnostic work depends on the knowledge and microscopy skills of the microbiologist.

Citation: Garcia L, Paltridge G, Shimizu R. 2015. General Approaches for Detection and Identification of Parasites, p 2317-2337. In Jorgensen J, Pfaller M, Carroll K, Funke G, Landry M, Richter S, Warnock D (ed), Manual of Clinical Microbiology, Eleventh Edition. ASM Press, Washington, DC. doi: 10.1128/9781555817381.ch135
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FIGURE 1

Processing liquid stool for O&P examination. Either PVA or Schaudinn’s fixative can be used for the preparation of the permanent stained smear. Organism motility is seen when saline is used; iodine kills the organisms, so motility will no longer be visible. The use of ethyl acetate may remove the entire specimen and pull it into the layer of debris that will be discarded (liquid specimen normally contains mucus); centrifuge at 500 × for 10 min (normal centrifugation time), but do not use ethyl acetate in the procedure. In general, laboratories have switched to nonmercury substitutes; the original Schaudinn’s fixative contains mercuric chloride. However, in some instances the term “Schaudinn’s fixative” is still used to describe not the original fixative but a formulation that is prepared with a copper or zinc base or other proprietary compounds. When fixatives are selected, it is important to know the contents in order to comply with disposal regulations. Reprinted from . doi:10.1128/9781555817381.ch135.f1

Citation: Garcia L, Paltridge G, Shimizu R. 2015. General Approaches for Detection and Identification of Parasites, p 2317-2337. In Jorgensen J, Pfaller M, Carroll K, Funke G, Landry M, Richter S, Warnock D (ed), Manual of Clinical Microbiology, Eleventh Edition. ASM Press, Washington, DC. doi: 10.1128/9781555817381.ch135
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Image of FIGURE 2
FIGURE 2

Processing preserved stool for O&P examination (two-vial collection kit). The formalin can be buffered or nonbuffered, depending on the laboratory protocol in use. Fixative prepared with mercuric chloride provides the best organism preservation. Alternatives are available, including zinc-based PVA, copper sulfate-based PVA, SAF, and the one of the single vial fixatives that requires no adhesive (PVA or albumin). Reprinted from . doi:10.1128/9781555817381.ch135.f2

Citation: Garcia L, Paltridge G, Shimizu R. 2015. General Approaches for Detection and Identification of Parasites, p 2317-2337. In Jorgensen J, Pfaller M, Carroll K, Funke G, Landry M, Richter S, Warnock D (ed), Manual of Clinical Microbiology, Eleventh Edition. ASM Press, Washington, DC. doi: 10.1128/9781555817381.ch135
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Image of FIGURE 3
FIGURE 3

Processing preserved stool for O&P examination (one-vial collection kit). There are single-vial collection systems for which the formulas are proprietary; however, many contain zinc sulfate as one of the key ingredients. With the exception of SAF, compatibility of these fixatives with immunoassay reagents is not always possible, particularly if the fixative contains PVA. Other good options include Unifix, Total-Fix (single-vial fixative with no mercury, formalin, or PVA) (Medical Chemical Corp.), or Z-PVA (Medical Chemical Corp.) with trichrome stain, as well as Ecofix with Ecostain (Meridian Biosciences, Cincinnati, OH). If the iron hematoxylin method containing the carbol fuchsin step is used, the coccidian oocysts will stain pink ( spp., , or ). Also, when using SAF, some recommend the use of an adhesive such as egg albumin to “glue” the fecal material onto the slide prior to staining. It is highly recommended that special stains be performed for the detection and identification of the coccidia (modified acid-fast stains) and the microsporidia (modified trichrome stains) from concentrated sediment to enhance organism recovery. Coccidian oocysts of can easily be detected in the concentration sediment wet mount; however, unless a very heavy infection is present, spp. oocysts may not be seen without special modified acid-fast stains. The small size of the microsporidian spores prevents identification without the use of special modified trichrome stains and microscopic examination with a 100× oil immersion objective. Reprinted from . doi:10.1128/9781555817381.ch135.f3

Citation: Garcia L, Paltridge G, Shimizu R. 2015. General Approaches for Detection and Identification of Parasites, p 2317-2337. In Jorgensen J, Pfaller M, Carroll K, Funke G, Landry M, Richter S, Warnock D (ed), Manual of Clinical Microbiology, Eleventh Edition. ASM Press, Washington, DC. doi: 10.1128/9781555817381.ch135
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References

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1. Garcia LS. 2007. Diagnostic Medical Parasitology, 5th ed. ASM Press, Washington, DC.
2. Hiatt RA,, Markell EK,, Ng E. 1995. How many stool examinations are necessary to detect pathogenic intestinal protozoa? Am J Trop Med Hyg 53:3639.
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5. National Committee for Clinical Laboratory Standards. 2000. Laboratory Diagnosis of Blood-Borne Parasitic Diseases. Approved Guideline M15-A. National Committee for Clinical Laboratory Standards, Villanova, PA.
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15. National Committee for Clinical Laboratory Standards. 1997. Procedures for the Recovery and Identification of Parasites from the Intestinal Tract. Approved Guideline M28-A. National Committee for Clinical Laboratory Standards, Villanova, PA.
16. Nielsen CK,, Ward LA. 1999. Enhanced detection of Cryptosporidium parvum in the acid-fast stain. J Vet Diagn Investig 11:567569.
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Tables

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TABLE 1

Diagnostic characteristics of organisms in wet mounts

Citation: Garcia L, Paltridge G, Shimizu R. 2015. General Approaches for Detection and Identification of Parasites, p 2317-2337. In Jorgensen J, Pfaller M, Carroll K, Funke G, Landry M, Richter S, Warnock D (ed), Manual of Clinical Microbiology, Eleventh Edition. ASM Press, Washington, DC. doi: 10.1128/9781555817381.ch135
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TABLE 2

Identification of helminth eggs

Citation: Garcia L, Paltridge G, Shimizu R. 2015. General Approaches for Detection and Identification of Parasites, p 2317-2337. In Jorgensen J, Pfaller M, Carroll K, Funke G, Landry M, Richter S, Warnock D (ed), Manual of Clinical Microbiology, Eleventh Edition. ASM Press, Washington, DC. doi: 10.1128/9781555817381.ch135
Generic image for table
TABLE 3

Diagnostic characteristics of organisms in permanent stained smears

Citation: Garcia L, Paltridge G, Shimizu R. 2015. General Approaches for Detection and Identification of Parasites, p 2317-2337. In Jorgensen J, Pfaller M, Carroll K, Funke G, Landry M, Richter S, Warnock D (ed), Manual of Clinical Microbiology, Eleventh Edition. ASM Press, Washington, DC. doi: 10.1128/9781555817381.ch135
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TABLE 4

Key to identification of intestinal amebae (permanent stained smear)

Citation: Garcia L, Paltridge G, Shimizu R. 2015. General Approaches for Detection and Identification of Parasites, p 2317-2337. In Jorgensen J, Pfaller M, Carroll K, Funke G, Landry M, Richter S, Warnock D (ed), Manual of Clinical Microbiology, Eleventh Edition. ASM Press, Washington, DC. doi: 10.1128/9781555817381.ch135
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TABLE 5

Key to identification of intestinal flagellates

Citation: Garcia L, Paltridge G, Shimizu R. 2015. General Approaches for Detection and Identification of Parasites, p 2317-2337. In Jorgensen J, Pfaller M, Carroll K, Funke G, Landry M, Richter S, Warnock D (ed), Manual of Clinical Microbiology, Eleventh Edition. ASM Press, Washington, DC. doi: 10.1128/9781555817381.ch135
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TABLE 6

Commercially available immunoassays for detection of intestinal parasites

Citation: Garcia L, Paltridge G, Shimizu R. 2015. General Approaches for Detection and Identification of Parasites, p 2317-2337. In Jorgensen J, Pfaller M, Carroll K, Funke G, Landry M, Richter S, Warnock D (ed), Manual of Clinical Microbiology, Eleventh Edition. ASM Press, Washington, DC. doi: 10.1128/9781555817381.ch135
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TABLE 7

Additional helminth recovery and identification techniques (other than O&P examination)

Citation: Garcia L, Paltridge G, Shimizu R. 2015. General Approaches for Detection and Identification of Parasites, p 2317-2337. In Jorgensen J, Pfaller M, Carroll K, Funke G, Landry M, Richter S, Warnock D (ed), Manual of Clinical Microbiology, Eleventh Edition. ASM Press, Washington, DC. doi: 10.1128/9781555817381.ch135
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TABLE 8

Recovery of parasites from other intestinal tract specimens

Citation: Garcia L, Paltridge G, Shimizu R. 2015. General Approaches for Detection and Identification of Parasites, p 2317-2337. In Jorgensen J, Pfaller M, Carroll K, Funke G, Landry M, Richter S, Warnock D (ed), Manual of Clinical Microbiology, Eleventh Edition. ASM Press, Washington, DC. doi: 10.1128/9781555817381.ch135
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TABLE 9

Detection of urogenital parasites

Citation: Garcia L, Paltridge G, Shimizu R. 2015. General Approaches for Detection and Identification of Parasites, p 2317-2337. In Jorgensen J, Pfaller M, Carroll K, Funke G, Landry M, Richter S, Warnock D (ed), Manual of Clinical Microbiology, Eleventh Edition. ASM Press, Washington, DC. doi: 10.1128/9781555817381.ch135
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TABLE 10

Specimen, possible parasite recovered, and appropriate tests (other than intestinal tract)

Citation: Garcia L, Paltridge G, Shimizu R. 2015. General Approaches for Detection and Identification of Parasites, p 2317-2337. In Jorgensen J, Pfaller M, Carroll K, Funke G, Landry M, Richter S, Warnock D (ed), Manual of Clinical Microbiology, Eleventh Edition. ASM Press, Washington, DC. doi: 10.1128/9781555817381.ch135
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TABLE 11

Techniques for the recovery and identification of blood parasites (EDTA or heparin) ( )

Citation: Garcia L, Paltridge G, Shimizu R. 2015. General Approaches for Detection and Identification of Parasites, p 2317-2337. In Jorgensen J, Pfaller M, Carroll K, Funke G, Landry M, Richter S, Warnock D (ed), Manual of Clinical Microbiology, Eleventh Edition. ASM Press, Washington, DC. doi: 10.1128/9781555817381.ch135

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