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Chapter 130 : Specimen Collection, Transport, and Processing: Parasitology

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Abstract:

This chapter deals with various specimen collection, transport, and processing methods for detection of parasites. Various specimen collection methods are available for specimens suspected of containing parasites or parasitic elements. The most common specimen submitted to the diagnostic laboratory is the stool specimen, and the most commonly performed procedure in parasitology is the ova and parasite (O&P) examination, which is composed of three separate protocols: the direct wet mount, the concentration, and the permanent stained smear. The examination of aspirated material for the diagnosis of parasitic infections may be extremely valuable, particularly when routine testing methods have failed to demonstrate the organisms. These specimens should be transported to the laboratory immediately after collection. Detection of parasites in tissue depends in part on specimen collection and on having sufficient material to perform the recommended diagnostic procedures. The nucleic acid-based diagnostic tests have been developed for almost all species of parasites. The main reason for the minor role of diagnostic PCR in parasitology is the fact that many parasite stages can be adequately diagnosed using established, more traditional techniques that are generally less expensive than PCR and technically less demanding. Diagnostic PCR may become more widespread when simple, fully standardized test kits are available and costs are reduced through the implementation of pre- and post-PCR automated techniques. Furthermore, the possibilities to not only detect and identify but also quantify organisms and determine their genotypes by analyzing the diagnostic PCR product extend the diagnostic power of PCR.

Citation: Shimizu R, Grimm F, Garcia L, Deplazes P. 2011. Specimen Collection, Transport, and Processing: Parasitology, p 2047-2063. In Versalovic J, Carroll K, Funke G, Jorgensen J, Landry M, Warnock D (ed), Manual of Clinical Microbiology, 10th Edition. ASM Press, Washington, DC. doi: 10.1128/9781555816728.ch130

Key Concept Ranking

Parasitic Diseases
0.50165206
Chagas' Disease
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Indirect Immunofluorescence Assay
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References

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Tables

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TABLE 1

Body sites and possible parasites recovered

Parasites include trophozoites, cysts, oocysts, spores, adults, larvae, eggs, and amastigote and trypomastigote stages. This table does not include every possible parasite that could be found in a particular body site. However, the most likely organisms have been listed.

Disseminated in severely immunosuppressed individuals.

Citation: Shimizu R, Grimm F, Garcia L, Deplazes P. 2011. Specimen Collection, Transport, and Processing: Parasitology, p 2047-2063. In Versalovic J, Carroll K, Funke G, Jorgensen J, Landry M, Warnock D (ed), Manual of Clinical Microbiology, 10th Edition. ASM Press, Washington, DC. doi: 10.1128/9781555816728.ch130
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TABLE 2

Commercially available kits for immunodetection of parasitic organisms or antigens in stool samples

A number of the kits are manufactured by a single manufacturer but labeled under different company names; consequently, some of the data for sensitivity and specificity may be identical to those of kits produced under another name or by another company.

EIA, enzyme immunoassay; DFA, direct fluorescent antibody; IC, immunochromatography.

URLs are given only the first time the company name appears in the table.

Citation: Shimizu R, Grimm F, Garcia L, Deplazes P. 2011. Specimen Collection, Transport, and Processing: Parasitology, p 2047-2063. In Versalovic J, Carroll K, Funke G, Jorgensen J, Landry M, Warnock D (ed), Manual of Clinical Microbiology, 10th Edition. ASM Press, Washington, DC. doi: 10.1128/9781555816728.ch130
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TABLE 3

Commercially available test kits for immunodetection or molecular detection of parasitic organisms or antigens in serum, plasma, blood, or vaginal discharge

Citation: Shimizu R, Grimm F, Garcia L, Deplazes P. 2011. Specimen Collection, Transport, and Processing: Parasitology, p 2047-2063. In Versalovic J, Carroll K, Funke G, Jorgensen J, Landry M, Warnock D (ed), Manual of Clinical Microbiology, 10th Edition. ASM Press, Washington, DC. doi: 10.1128/9781555816728.ch130
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TABLE 4

Commercially available kits or antigens for immunodetection of specific serum antibodies

Bordier Affinity Products, Chatanerie 2, CH-1023 Crissier, Switzerland (http://www.bordier.ch); Hemagen, 34–40 Bear Hill Rd., Waltham, MA 02154 (http:// www.hemagen.com); Immunetics, 380 Green St., Cambridge, MA 02139 (http://www.immunetics.com); InBios, 562 1st Ave. South, Suite 600, Seattle, WA 98104 (http://www.inbios.com); IVD Research, 5909 Sea Lion Place, Suite D, Carlsbad, CA 92008 (http://ivd@ivdresearch.com), bioMérieux SA, F-69280 Marcy l'Etoile, France (http://bioMerieux-diagnostics.com); Millipore Coporation, 290 Concord Rd., Billerica, MA (http://millipore.com); NovaTec, Immunodiagnostica GmbH, Waldstrasse 23 a6, 63128 Dietzenbach, Germany (http://www.novatec-id.com).

Abbreviations: EIA, enzyme immunoassay; Rapid, rapid immunochromatographic (some are dipstick, cartridge, or other rapid test formats).

Citation: Shimizu R, Grimm F, Garcia L, Deplazes P. 2011. Specimen Collection, Transport, and Processing: Parasitology, p 2047-2063. In Versalovic J, Carroll K, Funke G, Jorgensen J, Landry M, Warnock D (ed), Manual of Clinical Microbiology, 10th Edition. ASM Press, Washington, DC. doi: 10.1128/9781555816728.ch130
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TABLE 5

Specimen preparation and procedures, recommended stain(s) and relevant parasites, and additional information

CSF, cerebrospinal fluid; EIA, enzyme immunoassay; EITB, enzyme-linked immunoelectrotransfer blot (Western blot); EM, electron microscopy; FA, fluorescent antibody; GAE, granulomatous amebic encephalitis; GI, gastrointestinal; H & E, hematoxylin and eosin; IFA, indirect immunofluorescence assay; PAM, primary amebic encephalitis; PAS, periodic acid-Schiff stain; PBS, phosphate-buffered saline; QBC, quantitative buffy coat.

Many parasites or parasite stages may be detected in standard histologic sections of tissue material. However, species identification is difficult and additional examinations may be required. Usually, these techniques are not considered first-line methods. Additional methods like EM are carried out only by specialized laboratories and are not available for standard diagnostic purposes. EM examination for species identification has largely been replaced by PCR.

Material/specimens suitable for PCR: native (unfixed), in saline, PBS (ethanol), or frozen; avoid formalin.

Citation: Shimizu R, Grimm F, Garcia L, Deplazes P. 2011. Specimen Collection, Transport, and Processing: Parasitology, p 2047-2063. In Versalovic J, Carroll K, Funke G, Jorgensen J, Landry M, Warnock D (ed), Manual of Clinical Microbiology, 10th Edition. ASM Press, Washington, DC. doi: 10.1128/9781555816728.ch130
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TABLE 6

Fecal specimens for parasites: options for collection and processing

See key references and .

It is difficult to recognize an early outbreak situation where screening of all specimens for either , spp., or both may be relevant. If it appears that an outbreak is in the early stages, then performing the immunoassays on request can be changed to screening all stools.

Citation: Shimizu R, Grimm F, Garcia L, Deplazes P. 2011. Specimen Collection, Transport, and Processing: Parasitology, p 2047-2063. In Versalovic J, Carroll K, Funke G, Jorgensen J, Landry M, Warnock D (ed), Manual of Clinical Microbiology, 10th Edition. ASM Press, Washington, DC. doi: 10.1128/9781555816728.ch130
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TABLE 7

Approaches to stool parasitology: test ordering

Modified from reference .

Depending on the particular immunoassay kit used, various single or multiple organisms may be included. Selection of a particular kit depends on many variables: clinical relevance, cost, ease of performance, training, personnel availability, number of test orders, training of physician clients, sensitivity, specificity, equipment, time to result, etc. Very few laboratories handle this type of testing in exactly the same way. Many options are clinically relevant and acceptable for patient care.

Two stool specimens should be tested using an immunoassay in order to rule out an infection with fecal immunoassays may also be negative in cases with a low parasite load.

Citation: Shimizu R, Grimm F, Garcia L, Deplazes P. 2011. Specimen Collection, Transport, and Processing: Parasitology, p 2047-2063. In Versalovic J, Carroll K, Funke G, Jorgensen J, Landry M, Warnock D (ed), Manual of Clinical Microbiology, 10th Edition. ASM Press, Washington, DC. doi: 10.1128/9781555816728.ch130
Generic image for table
TABLE 8

Fecal preservatives: pros and cons

Modified acid-fast and modified trichrome stains.

Citation: Shimizu R, Grimm F, Garcia L, Deplazes P. 2011. Specimen Collection, Transport, and Processing: Parasitology, p 2047-2063. In Versalovic J, Carroll K, Funke G, Jorgensen J, Landry M, Warnock D (ed), Manual of Clinical Microbiology, 10th Edition. ASM Press, Washington, DC. doi: 10.1128/9781555816728.ch130

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