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Chapter 131 : Reagents, Stains, and Media: Parasitology

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Abstract:

The evaluation of clinical specimens for ova and parasites in the clinical laboratory can involve the use of direct macroscopic examination of the specimen and microscopic examination of fresh and preserved specimens, as well as culture for some parasitic organisms. These examinations necessitate the use of a variety of stains, reagents, and culture media, the most common of which are discussed in this chapter. Formalin has always been used in parasitology as an allpurpose preservative and in concentration procedures. The accompanying compound with the polyvinyl alcohol (PVA), specifically, mercuric chloride, zinc sulfate, or cupric sulfate, acts as the preservative and allows fixation of protozoan cysts and trophozoites for use with trichrome or iron hematoxylin stains for permanent smears. Specimens treated with Zn-PVA may also be stained with trichrome or iron hematoxylin stains for permanent smears. Specimens treated with Cu-PVA may also be stained with trichrome or iron hematoxylin stains for permanent smears. Examination of blood films for parasites includes the use of two common stains, the Giemsa stain and Wright's stain, both derivatives of the original Romanowsky stain. These stains are very similar, differing primarily in that no fixative is included in the Giemsa stain and the blood film must be fixed with absolute methanol prior to staining. In a basic staining procedure, thick films must be laked in distilled water or treated with saponin prior to performance of the staining procedure.

Citation: Linscott A, Sharp S. 2011. Reagents, Stains, and Media: Parasitology, p 2064-2070. 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.ch131

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References

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Tables

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TABLE 1

Preservatives used in diagnostic parasitology (intestinal tract specimens)

PVA (plastic powder used as “glue” to attach stool onto the glass slide/no fixation properties per se) and Schaudinn's fixative (mercuric chloride base) are still considered to be the “gold standard” against which all other fixatives are evaluated for organism morphology after permanent staining. Additional fixatives prepared with nonmercuric chloride-based compounds are continuing to be developed and tested.

This modification uses a copper sulfate base rather than mercuric chloride.

This modification uses a zinc base rather than mercuric chloride and apparently works well with both trichrome and iron hematoxylin stains.

These modifications use a combination of ingredients (including zinc) but are prepared from proprietary formulas. The aim is to provide a fixative that can be used for the fecal concentration, permanent stained smear, and available immunoassays for and spp. Some of these fixatives are now available commercially (check suppliers such as Medical Chemical Corp. and Meridian Bioscience). Testing for and/or the / group still require fresh or frozen specimens.

This stain can be used in place of trichrome for staining fecal smears preserved with MIF, PVA, or SAF methods. It is available commercially.

EIA, enzyme immunoassay; FA, fluorescent antibody; rapid, cartridge (membrane flow/immunochromatographic method).

Citation: Linscott A, Sharp S. 2011. Reagents, Stains, and Media: Parasitology, p 2064-2070. 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.ch131
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TABLE 2

Stains used for parasitic identification

Citation: Linscott A, Sharp S. 2011. Reagents, Stains, and Media: Parasitology, p 2064-2070. 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.ch131
Generic image for table
TABLE 3

Media used for cultivation of parasites

Citation: Linscott A, Sharp S. 2011. Reagents, Stains, and Media: Parasitology, p 2064-2070. 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.ch131

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