regardless of the suspected organism. The acid-fast trichrome procedure presented here also recommended the use of centrifuged fecal specimens.
5. Avoid the use of wet-gauze filtration (an old, standardized method of filtering stool prior to centrifugation) with too many layers of gauze that may trap organisms and not allow them to flow into the fluid to be concentrated. It is recommended that no more than two layers of gauze be used. Another option is to use the commercially available concentration systems that use metal or plastic screens for filtration.
References
1. California State Department of Public Health Microbial Disease Laboratory. 1979. PARA-1A Revised. California State Department of Public Health, Sacramento.
2. Clinical and Laboratory Standards Institute. 2005. Procedures for the Recovery and Identification of Parasites from the Intestinal Tract. Approved guideline M28-A2. Clinical and Laboratory Standards Institute, Villanova, PA.
3. Faust EC, D’Antoni JS, Odom V, Miller MF, Peres C, Sawitz W, Thomen LF, Tobie J, Walker JH. 1938. A critical study of clinical laboratory technics for the diagnosis of protozoan cysts and helminth eggs in feces. Am J Trop Med 18:169–183.
4. Garcia L, Brewer T, Bruckner D. 1979. A comparison of the formalin-ether concentration and trichrome-stained smear methods for the recovery and identification of intestinal protozoa. Am J Med Technol 45:932–935. PMID 92195
5. Garcia LS. 1990. Laboratory methods for diagnosis of parasitic infections, p 776–861. In Baron EJ, Finegold SM (ed), Bailey & Scott’s Diagnostic Microbiology, 8th ed. The C. V. Mosby Co., St. Louis, MO.
6. Gleason NN, Healy GR. 1965. Modification and evaluation of Kohn’s one-step staining technic for intestinal protozoa in feces or tissue. Am J Clin Pathol 43:494–496. PMID 14289880
7. Gomori G. 1950. A rapid one-step trichrome stain. Am J Clin Pathol 20:661–663. PMID 15432364
8. Kellogg JA, Elder CJ. 1999. Justification for use of a single trichrome stain as the sole means for routine detection of intestinal parasites in concentrated stool specimens. J Clin Microbiol 37:835–837. PMID 9986870
9. Kohn J. 1960. A one stage permanent staining method for fecal protozoa. Dapim Refuiim Med Q Isr 19:160–161.
10. Lawson LL, Bailey JW, Beeching NJ, Gurgel RG, Cuevas LE. 2004. The stool examination reports amoeba cysts: should you treat in the face of over diagnosis and lack of specificity of light microscopy? Trop Doct 34:28–30. PMID 14959971
11. Levine JA, Estevez EG. 1983. Method for concentration of parasites from small amounts of feces. J Clin Microbiol 18:786–788. PMID 6630461
12. Markell EK, Voge M. 1981. Medical Parasitology, 5th ed. The W. B. Saunders Co., Philadelphia, PA.
13. Melvin DM, Brooke MM. 1982. Laboratory Procedures for the Diagnosis of Intestinal Parasites, 3rd ed. U.S. Department of Health, Education, and Welfare publication (CDC) 82-8282. Government Printing Office, Washington, DC.
14. Miller JM. 1991. Quality control of media, reagents, and stains, p. 1203–1225. In Balows A, Hausler WJ Jr, Herrmann KL, Isenberg HD, Shadomy HJ (ed.), Manual of Clinical Microbiology, 5th ed. American Society for Microbiology, Washington, DC.
15. Nair C. 1953. Rapid staining of intestinal amoebae on wet mounts. Nature (London) 172:1051.
16. Ritchie L. 1948. An ether sedimentation technique for routine stool examinations. Bull US Army Med Dept 8:326. PMID 18911509
17. Shoaib S, Hafiz A, Tauheed S. 2002. Role of trichrome staining techniques in the diagnosis of intestinal parasitic infections. J Pak Med Assoc 52:152–154. PMID 12174479
18. Smith JW, Bartlett MS. 1985. Diagnostic parasitology: introduction and methods, p. 595–611. In Lennette EH, Balows A, Hausler WJ Jr, Shadomy HJ (ed.), Manual of Clinical Microbiology, 4th ed. American Society for Microbiology, Washington, DC.
19. Spencer FM, Monroe LS. 1976. The Color Atlas of Intestinal Parasites, 2nd ed. Charles C Thomas, Publisher, Springfield, IL.
20. Tompkins VN, Miller JK. 1947. Staining intestinal protozoa with iron-hematoxylin-phosphotungstic acid. Am J Clin Pathol 17:755–758. PMID 20265758
21. Truant AL, Elliott SH, Kelley MY, Smith JH. 1981. Comparison of formalin-ethyl ether sedimentation, formalin-ethyl acetate sedimentation, and zinc sulfate flotation techniques for detection of intestinal parasites. J Clin Microbiol 13:882–884. PMID 7240400
22. Wheatley W. 1951. A rapid staining procedure for intestinal amoebae and flagellates. Am J Clin Pathol 21:990–991. PMID 14885115
23. Neimeister R, Logan A, Gerber B, Egleton J, Kleger B. 1987. Hemo-De as substitute for ethyl acetate in formalin-ethyl acetate concentration technique. J Clin Microbiol 25:425–426. PMID 3818930
24. Higgins GV. 1988. Iodine-trichrome staining technique: a replacement of iodine solution in fecal examination. Lab Med 19:824–825.
25. Hanson KL, Cartwright CP. 2001. Use of an enzyme immunoassay does not eliminate the need to analyze stool specimens for sensitive detection of Giardia lamblia. J Clin Microbiol 39:474–477. PMID 11158092
26. Morimoto N, Komatsu C, Nishida M, Sugiura T. 2001. Detection of Giardia lamblia cysts in non-fixed long-term stored human feces by direct immunofluorescence assay. Jpn J Infect Dis 54:72–74. PMID 11427745
27. Fedorko DP, Williams EC, Nelson NA, Mazyck TD, Hanson KL, Cartwright CP. 2000. Performance of Para-Pak Ultra ECOFIX compared with Para-Pak Ultra formalin/mercuric chloride-based polyvinyl alcohol for concentration and permanent stained smears of stool parasites. Diagn Microbiol Infect Dis 37:37–39. PMID 10794938
28. Garcia LS, Shimizu RY. 1998. Evaluation of intestinal protozoan morphology in human fecal specimens preserved in EcoFix: comparison of Wheatley’s trichrome stain and EcoStain. J Clin Microbiol 36:1974–1976. PMID 9650947
29. Garcia LS, Shimizu RY, Shum AC, Bruckner DA. 1993. Evaluation of intestinal protozoan morphology in polyvinyl alcohol preservative: comparison of zinc sulfate- and mercuric chloride-based compounds for use in Schaudinn’s fixative. J Clin Microbiol 31: 307–310.
30. Ignatius R, Lehmann M, Miksits K, Regnath T, Arvand M, Engelmann E, Futh U, Hahn H, Wagner J. 1997. A new acid-fast trichrome stain for simultaneous detection of Cryptosporidium parvum and microsporidial species in stool specimens. J Clin Microbiol 35:446–449. PMID 9003613
31.
