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Chapter 5 : Specific Test Procedures and Algorithms
Editor:
Lynne S. Garcia1
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1: LSG & Associates, Santa Monica, California
Content Type: Reference
Publication Year:
2009
Category: Clinical Microbiology
Book DOI:
10.1128/9781555815967
Chapter DOI:
10.1128/9781555815967.ch5
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Abstract:
This section focuses on microscopic techniques and calibration of the microscope and algorithms for the ova and parasite (O&P) examination. Direct wet fecal smear, sedimentation concentration, flotation concentration methods used for the results and laboratory reports and procedure reminders and limitations are briefly discussed. Specialized stains for Coccidia and Microsporidia are discussed using different staining methods. Modified trichrome stain for Microsporidia is reviewed for the clinical assay. Antigen-based fecal immunoassays have several significant advantages over other methods currently used for diagnosis of amebiasis. Different stains are analyzed for the parasites examination. Several concentration techniques such as buffy coat concentration, knott concentration and membrane filtration concentration are discussed. The section also provides a brief outline of the procedure for processing fresh stool for the O&P examination.
Citation:
Garcia L.
2009.
Specific Test Procedures and Algorithms,
p 87-217.
In
Practical Guide to Diagnostic Parasitology, Second Edition.
ASM Press, Washington, DC.
doi: 10.1128/9781555815967.ch5
Figures
Specific Test Procedures and Algorithms
Citation:
Garcia L.
2009.
Specific Test Procedures and Algorithms,
p 87-217.
In
Practical Guide to Diagnostic Parasitology, Second Edition.
ASM Press, Washington, DC.
doi: 10.1128/9781555815967.ch5
/content/10.1128/9781555815967.ch05.ch05fig01
Figure 5.1
Ocular micrometer, top scale; stage micrometer, bottom scale. Reprinted from Diagnostic Medical Parasitology, 5th ed.
10.1128/9781555815967/ch5fig1_thmb.gif
10.1128/9781555815967/ch5fig1.gif
Figure 5.1
Ocular micrometer, top scale; stage micrometer, bottom scale. Reprinted from Diagnostic Medical Parasitology, 5th ed.
Citation:
Garcia L.
2009.
Specific Test Procedures and Algorithms,
p 87-217.
In
Practical Guide to Diagnostic Parasitology, Second Edition.
ASM Press, Washington, DC.
doi: 10.1128/9781555815967.ch5
/content/10.1128/9781555815967.ch05.ch05eq01
Untitled
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Citation:
Garcia L.
2009.
Specific Test Procedures and Algorithms,
p 87-217.
In
Practical Guide to Diagnostic Parasitology, Second Edition.
ASM Press, Washington, DC.
doi: 10.1128/9781555815967.ch5
/content/10.1128/9781555815967.ch05.ch05eq02
Untitled
10.1128/9781555815967/ch5equ2_thmb.gif
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Untitled
Citation:
Garcia L.
2009.
Specific Test Procedures and Algorithms,
p 87-217.
In
Practical Guide to Diagnostic Parasitology, Second Edition.
ASM Press, Washington, DC.
doi: 10.1128/9781555815967.ch5
/content/10.1128/9781555815967.ch05.ch05eq03
Untitled
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Untitled
Citation:
Garcia L.
2009.
Specific Test Procedures and Algorithms,
p 87-217.
In
Practical Guide to Diagnostic Parasitology, Second Edition.
ASM Press, Washington, DC.
doi: 10.1128/9781555815967.ch5
/content/10.1128/9781555815967.ch05.ch05eq04
Untitled
10.1128/9781555815967/ch5equ4_thmb.gif
10.1128/9781555815967/ch5equ4.gif
Untitled
Citation:
Garcia L.
2009.
Specific Test Procedures and Algorithms,
p 87-217.
In
Practical Guide to Diagnostic Parasitology, Second Edition.
ASM Press, Washington, DC.
doi: 10.1128/9781555815967.ch5
/content/10.1128/9781555815967.ch05.ch05fig02
Figure 5.2
Method of scanning a direct wet film preparation with the 10× objective. (Illustration by Nobuko Kitamura.) Reprinted from Diagnostic Medical Parasitology, 5th ed.
10.1128/9781555815967/ch5fig2_thmb.gif
10.1128/9781555815967/ch5fig2.gif
Figure 5.2
Method of scanning a direct wet film preparation with the 10× objective. (Illustration by Nobuko Kitamura.) Reprinted from Diagnostic Medical Parasitology, 5th ed.
Citation:
Garcia L.
2009.
Specific Test Procedures and Algorithms,
p 87-217.
In
Practical Guide to Diagnostic Parasitology, Second Edition.
ASM Press, Washington, DC.
doi: 10.1128/9781555815967.ch5
/content/10.1128/9781555815967.ch05.ch05fig03
Figure 5.3
Fecal concentration procedures: various layers seen in tubes after centrifugation. (A) Formalin-ether (or ethyl acetate). (B) Zinc sulfate (the surface film should be within 2 to 3 mm of the tube rim). (Illustration by Sharon Belkin.)
10.1128/9781555815967/ch5fig3_thmb.gif
10.1128/9781555815967/ch5fig3.gif
Figure 5.3
Fecal concentration procedures: various layers seen in tubes after centrifugation. (A) Formalin-ether (or ethyl acetate). (B) Zinc sulfate (the surface film should be within 2 to 3 mm of the tube rim). (Illustration by Sharon Belkin.)
Citation:
Garcia L.
2009.
Specific Test Procedures and Algorithms,
p 87-217.
In
Practical Guide to Diagnostic Parasitology, Second Edition.
ASM Press, Washington, DC.
doi: 10.1128/9781555815967.ch5
/content/10.1128/9781555815967.ch05.ch05fig04
Figure 5.4
Method used to remove surface film in the zinc sulfate flotation concentration procedure. (A) A wire loop is gently placed on (not under) the surface film. (B) The loop is then placed on a glass slide. (Illustration by Nobuko Kitamura.) Reprinted from Diagnostic Medical Parasitology, 5th ed.
10.1128/9781555815967/ch5fig4_thmb.gif
10.1128/9781555815967/ch5fig4.gif
Figure 5.4
Method used to remove surface film in the zinc sulfate flotation concentration procedure. (A) A wire loop is gently placed on (not under) the surface film. (B) The loop is then placed on a glass slide. (Illustration by Nobuko Kitamura.) Reprinted from Diagnostic Medical Parasitology, 5th ed.
Citation:
Garcia L.
2009.
Specific Test Procedures and Algorithms,
p 87-217.
In
Practical Guide to Diagnostic Parasitology, Second Edition.
ASM Press, Washington, DC.
doi: 10.1128/9781555815967.ch5
/content/10.1128/9781555815967.ch05.ch05fig05
Figure 5.5
Culture methods for the recovery of larval-stage nematodes; Harada-Mori tube method and petri dish culture method. (Illustration by Nobuko Kitamura.) Reprinted from Diagnostic Medical Parasitology, 5th ed.
10.1128/9781555815967/ch5fig5_thmb.gif
10.1128/9781555815967/ch5fig5.gif
Figure 5.5
Culture methods for the recovery of larval-stage nematodes; Harada-Mori tube method and petri dish culture method. (Illustration by Nobuko Kitamura.) Reprinted from Diagnostic Medical Parasitology, 5th ed.
Citation:
Garcia L.
2009.
Specific Test Procedures and Algorithms,
p 87-217.
In
Practical Guide to Diagnostic Parasitology, Second Edition.
ASM Press, Washington, DC.
doi: 10.1128/9781555815967.ch5
/content/10.1128/9781555815967.ch05.ch05fig06
Figure 5.6
Baermann apparatus. (Illustration by Nobuko Kitamura.) Reprinted from Diagnostic Medical Parasitology, 5th ed.
10.1128/9781555815967/ch5fig6_thmb.gif
10.1128/9781555815967/ch5fig6.gif
Figure 5.6
Baermann apparatus. (Illustration by Nobuko Kitamura.) Reprinted from Diagnostic Medical Parasitology, 5th ed.
Citation:
Garcia L.
2009.
Specific Test Procedures and Algorithms,
p 87-217.
In
Practical Guide to Diagnostic Parasitology, Second Edition.
ASM Press, Washington, DC.
doi: 10.1128/9781555815967.ch5
/content/10.1128/9781555815967.ch05.ch05fig07
Figure 5.7
Agar culture method for Strongyloides stercoralis. (1) Agar plates are prepared; (2) agar is dried for 4 to 5 days on the bench top; (3) plates are stored in plastic bags; (4) fresh stool is submitted to the laboratory; (5) approximately 2 g of stool is placed onto an agar plate; (6) the plate is sealed with tape; (7) the culture plate is incubated at 26 to 33°C for 2 days; (8) the plate is examined microscopically for the presence of tracks (bacteria carried over the agar by migrating larvae); (9) 10% formalin is placed onto the agar through a hole made in the plastic by hot forceps; (10) material from the agar plate is centrifuged; (11) the material is examined as a wet preparation for rhabditiform or filariform larvae (high dry power; magnification, ×400). (Illustration by Sharon Belkin.) Reprinted from Diagnostic Medical Parasitology, 5th ed.
10.1128/9781555815967/ch5fig7_thmb.gif
10.1128/9781555815967/ch5fig7.gif
Figure 5.7
Agar culture method for Strongyloides stercoralis. (1) Agar plates are prepared; (2) agar is dried for 4 to 5 days on the bench top; (3) plates are stored in plastic bags; (4) fresh stool is submitted to the laboratory; (5) approximately 2 g of stool is placed onto an agar plate; (6) the plate is sealed with tape; (7) the culture plate is incubated at 26 to 33°C for 2 days; (8) the plate is examined microscopically for the presence of tracks (bacteria carried over the agar by migrating larvae); (9) 10% formalin is placed onto the agar through a hole made in the plastic by hot forceps; (10) material from the agar plate is centrifuged; (11) the material is examined as a wet preparation for rhabditiform or filariform larvae (high dry power; magnification, ×400). (Illustration by Sharon Belkin.) Reprinted from Diagnostic Medical Parasitology, 5th ed.
Citation:
Garcia L.
2009.
Specific Test Procedures and Algorithms,
p 87-217.
In
Practical Guide to Diagnostic Parasitology, Second Edition.
ASM Press, Washington, DC.
doi: 10.1128/9781555815967.ch5
/content/10.1128/9781555815967.ch05.ch05fig08
Figure 5.8
Method of thin blood film preparation. (A) Position of spreader slide; (B) well-prepared thin film. Arrows indicate the area of the slide used to observe accurate cell morphology. (Illustration by Nobuko Kitamura.) Reprinted from Diagnostic Medical Parasitology, 5th ed.
10.1128/9781555815967/ch5fig8_thmb.gif
10.1128/9781555815967/ch5fig8.gif
Figure 5.8
Method of thin blood film preparation. (A) Position of spreader slide; (B) well-prepared thin film. Arrows indicate the area of the slide used to observe accurate cell morphology. (Illustration by Nobuko Kitamura.) Reprinted from Diagnostic Medical Parasitology, 5th ed.
Citation:
Garcia L.
2009.
Specific Test Procedures and Algorithms,
p 87-217.
In
Practical Guide to Diagnostic Parasitology, Second Edition.
ASM Press, Washington, DC.
doi: 10.1128/9781555815967.ch5
/content/10.1128/9781555815967.ch05.ch05fig09
Figure 5.9
Method of thick-thin combination blood film preparation. (a) Position of a drop of EDTA-blood; (b) position of the applicator stick in contact with blood and glass slide; (c) rotation of the applicator stick; and (d) completed thick-thin combination blood film prior to staining. (Illustration by Sharon Belkin.) Reprinted from Diagnostic Medical Parasitology, 5th ed.
10.1128/9781555815967/ch5fig9_thmb.gif
10.1128/9781555815967/ch5fig9.gif
Figure 5.9
Method of thick-thin combination blood film preparation. (a) Position of a drop of EDTA-blood; (b) position of the applicator stick in contact with blood and glass slide; (c) rotation of the applicator stick; and (d) completed thick-thin combination blood film prior to staining. (Illustration by Sharon Belkin.) Reprinted from Diagnostic Medical Parasitology, 5th ed.
Citation:
Garcia L.
2009.
Specific Test Procedures and Algorithms,
p 87-217.
In
Practical Guide to Diagnostic Parasitology, Second Edition.
ASM Press, Washington, DC.
doi: 10.1128/9781555815967.ch5
/content/10.1128/9781555815967.ch05.ch05alg01
Algorithm 5.1
Procedure for processing fresh stool for the O&P examination
10.1128/9781555815967/201fig1_thmb.gif
10.1128/9781555815967/201fig1.gif
Algorithm 5.1
Procedure for processing fresh stool for the O&P examination
Citation:
Garcia L.
2009.
Specific Test Procedures and Algorithms,
p 87-217.
In
Practical Guide to Diagnostic Parasitology, Second Edition.
ASM Press, Washington, DC.
doi: 10.1128/9781555815967.ch5
/content/10.1128/9781555815967.ch05.ch05alg02
Algorithm 5.2
Procedure for processing liquid specimens for the O&P examination
10.1128/9781555815967/202fig1_thmb.gif
10.1128/9781555815967/202fig1.gif
Algorithm 5.2
Procedure for processing liquid specimens for the O&P examination
Citation:
Garcia L.
2009.
Specific Test Procedures and Algorithms,
p 87-217.
In
Practical Guide to Diagnostic Parasitology, Second Edition.
ASM Press, Washington, DC.
doi: 10.1128/9781555815967.ch5
/content/10.1128/9781555815967.ch05.ch05alg03
Algorithm 5.3
Procedure for processing preserved stool for the O&P examination—two-vial collection kit
10.1128/9781555815967/203fig1_thmb.gif
10.1128/9781555815967/203fig1.gif
Algorithm 5.3
Procedure for processing preserved stool for the O&P examination—two-vial collection kit
Citation:
Garcia L.
2009.
Specific Test Procedures and Algorithms,
p 87-217.
In
Practical Guide to Diagnostic Parasitology, Second Edition.
ASM Press, Washington, DC.
doi: 10.1128/9781555815967.ch5
/content/10.1128/9781555815967.ch05.ch05alg04
Algorithm 5.4
Procedure for processing SAF-preserved stool for the O&P examination
10.1128/9781555815967/204fig1_thmb.gif
10.1128/9781555815967/204fig1.gif
Algorithm 5.4
Procedure for processing SAF-preserved stool for the O&P examination
Citation:
Garcia L.
2009.
Specific Test Procedures and Algorithms,
p 87-217.
In
Practical Guide to Diagnostic Parasitology, Second Edition.
ASM Press, Washington, DC.
doi: 10.1128/9781555815967.ch5
/content/10.1128/9781555815967.ch05.ch05alg05
Algorithm 5.5
Use of various fixatives and their recommended stains
10.1128/9781555815967/205fig1_thmb.gif
10.1128/9781555815967/205fig1.gif
Algorithm 5.5
Use of various fixatives and their recommended stains
Citation:
Garcia L.
2009.
Specific Test Procedures and Algorithms,
p 87-217.
In
Practical Guide to Diagnostic Parasitology, Second Edition.
ASM Press, Washington, DC.
doi: 10.1128/9781555815967.ch5
/content/10.1128/9781555815967.ch05.ch05alg06
Algorithm 5.6
Ordering algorithm for laboratory examination for intestinal parasites
10.1128/9781555815967/206fig1_thmb.gif
10.1128/9781555815967/206fig1.gif
Algorithm 5.6
Ordering algorithm for laboratory examination for intestinal parasites
Citation:
Garcia L.
2009.
Specific Test Procedures and Algorithms,
p 87-217.
In
Practical Guide to Diagnostic Parasitology, Second Edition.
ASM Press, Washington, DC.
doi: 10.1128/9781555815967.ch5
/content/10.1128/9781555815967.ch05.ch05alg07
Algorithm 5.7
Procedure for processing blood specimens for examination
10.1128/9781555815967/208fig1_thmb.gif
10.1128/9781555815967/208fig1.gif
Algorithm 5.7
Procedure for processing blood specimens for examination
Citation:
Garcia L.
2009.
Specific Test Procedures and Algorithms,
p 87-217.
In
Practical Guide to Diagnostic Parasitology, Second Edition.
ASM Press, Washington, DC.
doi: 10.1128/9781555815967.ch5
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Tables
Specific Test Procedures and Algorithms
Citation:
Garcia L.
2009.
Specific Test Procedures and Algorithms,
p 87-217.
In
Practical Guide to Diagnostic Parasitology, Second Edition.
ASM Press, Washington, DC.
doi: 10.1128/9781555815967.ch5
/content/10.1128/9781555815967.ch05.ch05nt08
Untitled
Untitled
Citation:
Garcia L.
2009.
Specific Test Procedures and Algorithms,
p 87-217.
In
Practical Guide to Diagnostic Parasitology, Second Edition.
ASM Press, Washington, DC.
doi: 10.1128/9781555815967.ch5
/content/10.1128/9781555815967.ch05.ch05nt18
Untitled
Untitled
Citation:
Garcia L.
2009.
Specific Test Procedures and Algorithms,
p 87-217.
In
Practical Guide to Diagnostic Parasitology, Second Edition.
ASM Press, Washington, DC.
doi: 10.1128/9781555815967.ch5
/content/10.1128/9781555815967.ch05.ch05nt21
Untitled
Untitled
Citation:
Garcia L.
2009.
Specific Test Procedures and Algorithms,
p 87-217.
In
Practical Guide to Diagnostic Parasitology, Second Edition.
ASM Press, Washington, DC.
doi: 10.1128/9781555815967.ch5
/content/10.1128/9781555815967.ch05.ch05nt33
Untitled
Untitled
Citation:
Garcia L.
2009.
Specific Test Procedures and Algorithms,
p 87-217.
In
Practical Guide to Diagnostic Parasitology, Second Edition.
ASM Press, Washington, DC.
doi: 10.1128/9781555815967.ch5
/content/10.1128/9781555815967.ch05.ch05nt37
Untitled
Untitled
Citation:
Garcia L.
2009.
Specific Test Procedures and Algorithms,
p 87-217.
In
Practical Guide to Diagnostic Parasitology, Second Edition.
ASM Press, Washington, DC.
doi: 10.1128/9781555815967.ch5
/content/10.1128/9781555815967.ch05.ch05nt43
Untitled
Untitled
Citation:
Garcia L.
2009.
Specific Test Procedures and Algorithms,
p 87-217.
In
Practical Guide to Diagnostic Parasitology, Second Edition.
ASM Press, Washington, DC.
doi: 10.1128/9781555815967.ch5
/content/10.1128/9781555815967.ch05.ch05nt47
Untitled
Untitled
Citation:
Garcia L.
2009.
Specific Test Procedures and Algorithms,
p 87-217.
In
Practical Guide to Diagnostic Parasitology, Second Edition.
ASM Press, Washington, DC.
doi: 10.1128/9781555815967.ch5
/content/10.1128/9781555815967.ch05.ch05nt55
Untitled
Untitled
Citation:
Garcia L.
2009.
Specific Test Procedures and Algorithms,
p 87-217.
In
Practical Guide to Diagnostic Parasitology, Second Edition.
ASM Press, Washington, DC.
doi: 10.1128/9781555815967.ch5
/content/10.1128/9781555815967.ch05.ch05nt66
Untitled
Untitled
Citation:
Garcia L.
2009.
Specific Test Procedures and Algorithms,
p 87-217.
In
Practical Guide to Diagnostic Parasitology, Second Edition.
ASM Press, Washington, DC.
doi: 10.1128/9781555815967.ch5
/content/10.1128/9781555815967.ch05.ch05nt74
Untitled
Untitled
Citation:
Garcia L.
2009.
Specific Test Procedures and Algorithms,
p 87-217.
In
Practical Guide to Diagnostic Parasitology, Second Edition.
ASM Press, Washington, DC.
doi: 10.1128/9781555815967.ch5
/content/10.1128/9781555815967.ch05.ch05tab01
Table 5.1
Body site, specimen, and recommended stain(s)
a
Table 5.1
Body site, specimen, and recommended stain(s) a
Citation:
Garcia L.
2009.
Specific Test Procedures and Algorithms,
p 87-217.
In
Practical Guide to Diagnostic Parasitology, Second Edition.
ASM Press, Washington, DC.
doi: 10.1128/9781555815967.ch5
/content/10.1128/9781555815967.ch05.ch05tab02
Table 5.2
Approaches to stool parasitology: test ordering
Table 5.2
Approaches to stool parasitology: test ordering
Citation:
Garcia L.
2009.
Specific Test Procedures and Algorithms,
p 87-217.
In
Practical Guide to Diagnostic Parasitology, Second Edition.
ASM Press, Washington, DC.
doi: 10.1128/9781555815967.ch5
/content/10.1128/9781555815967.ch05.ch05tab03
Table 5.3
Laboratory test reports: optional comments
a,b
Table 5.3
Laboratory test reports: optional comments a,b
Citation:
Garcia L.
2009.
Specific Test Procedures and Algorithms,
p 87-217.
In
Practical Guide to Diagnostic Parasitology, Second Edition.
ASM Press, Washington, DC.
doi: 10.1128/9781555815967.ch5
/content/10.1128/9781555815967.ch05.ch05tab04
Table 5.4
Parasitemia determined from conventional light microscopy: clinical correlation
Table 5.4
Parasitemia determined from conventional light microscopy: clinical correlation
Citation:
Garcia L.
2009.
Specific Test Procedures and Algorithms,
p 87-217.
In
Practical Guide to Diagnostic Parasitology, Second Edition.
ASM Press, Washington, DC.
doi: 10.1128/9781555815967.ch5