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2013 PARASITOLOGY WORKSHOP LYNNE S. GARCIA, MS, FAAM, CLS, BLM Diagnostic Medical Parasitology Workshop 2013 UPDATE – QUIZ PART 7 ORGANISM IDENTIFICATIONS, KEY QUESTIONS SPONSORED BY MEDICAL CHEMICAL CORPORATION www.med-chem.com
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2013 PARASITOLOGY WORKSHOP LYNNE S. GARCIA, MS, FAAM, CLS, BLM Diagnostic Medical Parasitology Workshop 2013 UPDATE – QUIZ PART 7 ORGANISM IDENTIFICATIONS,

Dec 24, 2015

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Page 1: 2013 PARASITOLOGY WORKSHOP LYNNE S. GARCIA, MS, FAAM, CLS, BLM Diagnostic Medical Parasitology Workshop 2013 UPDATE – QUIZ PART 7 ORGANISM IDENTIFICATIONS,

2013 PARASITOLOGY WORKSHOP

LYNNE S. GARCIA, MS, FAAM, CLS, BLM

Diagnostic Medical Parasitology Workshop

2013 UPDATE – QUIZ PART 7 ORGANISM IDENTIFICATIONS, KEY QUESTIONS

SPONSORED BY

MEDICAL CHEMICAL CORPORATIONwww.med-chem.com

Page 2: 2013 PARASITOLOGY WORKSHOP LYNNE S. GARCIA, MS, FAAM, CLS, BLM Diagnostic Medical Parasitology Workshop 2013 UPDATE – QUIZ PART 7 ORGANISM IDENTIFICATIONS,

ORGANISM # 1 2

Duodenal Fluid, Modified Trichrome StainGeneral name is acceptable.

Page 3: 2013 PARASITOLOGY WORKSHOP LYNNE S. GARCIA, MS, FAAM, CLS, BLM Diagnostic Medical Parasitology Workshop 2013 UPDATE – QUIZ PART 7 ORGANISM IDENTIFICATIONS,

ORGANISM # 2

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Stool smear stained with Wheatley’s Trichrome stain.

Give genus, species, stage names

Page 4: 2013 PARASITOLOGY WORKSHOP LYNNE S. GARCIA, MS, FAAM, CLS, BLM Diagnostic Medical Parasitology Workshop 2013 UPDATE – QUIZ PART 7 ORGANISM IDENTIFICATIONS,

QUESTION 1

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Is it good laboratory practice to accept 2 (rather than 3) stools for the O&P examination? Why or why not?

Page 5: 2013 PARASITOLOGY WORKSHOP LYNNE S. GARCIA, MS, FAAM, CLS, BLM Diagnostic Medical Parasitology Workshop 2013 UPDATE – QUIZ PART 7 ORGANISM IDENTIFICATIONS,

ANSWER 1

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Three stool specimens is more accurate, but many laboratories feel organism recovery with 2 stools is acceptable. Percent recovery varies from ~80% to over 95% with the submission and examination of two stool specimens. A single stool specimen is only about 50% accurate. In one study, they found the following increases examining the third stool: Hiatt, et al. (Am. J. Trop. Med. Hyg. 53:36-9, 1995)

Yield increased 22.7%: Entamoeba histolyticaYield increased 11.3%: Giardia lambliaYield increased 31.1%: Dientamoeba fragilis

Page 6: 2013 PARASITOLOGY WORKSHOP LYNNE S. GARCIA, MS, FAAM, CLS, BLM Diagnostic Medical Parasitology Workshop 2013 UPDATE – QUIZ PART 7 ORGANISM IDENTIFICATIONS,

ORGANISM # 3

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Fluorescence immunoassay.What are the two structures?

Use genus, species names

Page 7: 2013 PARASITOLOGY WORKSHOP LYNNE S. GARCIA, MS, FAAM, CLS, BLM Diagnostic Medical Parasitology Workshop 2013 UPDATE – QUIZ PART 7 ORGANISM IDENTIFICATIONS,

ORGANISM # 47

Blood Stain

Name genus and species

What’s the clue?

Page 8: 2013 PARASITOLOGY WORKSHOP LYNNE S. GARCIA, MS, FAAM, CLS, BLM Diagnostic Medical Parasitology Workshop 2013 UPDATE – QUIZ PART 7 ORGANISM IDENTIFICATIONS,

QUESTION 2

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When selecting a single vial stool fixative, what questions should be asked?

What are Universal Fixatives?

Page 9: 2013 PARASITOLOGY WORKSHOP LYNNE S. GARCIA, MS, FAAM, CLS, BLM Diagnostic Medical Parasitology Workshop 2013 UPDATE – QUIZ PART 7 ORGANISM IDENTIFICATIONS,

ANSWER 2

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Can one perform a concentration, permanent stained smear, special stains for the coccidia and microsporidia, and/or fecal immunoassay procedures from the specimen received in that vial (Universal Fixative)? What stains work best with that particular fixative?

TOTAL-FIX contains NO mercury, NO formalin, and NO PVA.

Page 10: 2013 PARASITOLOGY WORKSHOP LYNNE S. GARCIA, MS, FAAM, CLS, BLM Diagnostic Medical Parasitology Workshop 2013 UPDATE – QUIZ PART 7 ORGANISM IDENTIFICATIONS,

ORGANISM # 5

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Stool smear stained with Wheatley’s Trichrome stain.

Name genus, species, and stage

Page 11: 2013 PARASITOLOGY WORKSHOP LYNNE S. GARCIA, MS, FAAM, CLS, BLM Diagnostic Medical Parasitology Workshop 2013 UPDATE – QUIZ PART 7 ORGANISM IDENTIFICATIONS,

ORGANISM # 611

Blood Stain;

Name Genus;

What is the structure within the circle called? Is it always seen?O

Page 12: 2013 PARASITOLOGY WORKSHOP LYNNE S. GARCIA, MS, FAAM, CLS, BLM Diagnostic Medical Parasitology Workshop 2013 UPDATE – QUIZ PART 7 ORGANISM IDENTIFICATIONS,

ORGANISM # 7

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Blood StainWhat is general term for structures?Name genus and species (bone marrow).

Page 13: 2013 PARASITOLOGY WORKSHOP LYNNE S. GARCIA, MS, FAAM, CLS, BLM Diagnostic Medical Parasitology Workshop 2013 UPDATE – QUIZ PART 7 ORGANISM IDENTIFICATIONS,

ORGANISM # 8

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Blood Stain; name genus, species

Page 14: 2013 PARASITOLOGY WORKSHOP LYNNE S. GARCIA, MS, FAAM, CLS, BLM Diagnostic Medical Parasitology Workshop 2013 UPDATE – QUIZ PART 7 ORGANISM IDENTIFICATIONS,

QUESTION 3

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What is PVA (stool adhesive)?

Does it work like albumin?

Does every fixative require one or the other?

Page 15: 2013 PARASITOLOGY WORKSHOP LYNNE S. GARCIA, MS, FAAM, CLS, BLM Diagnostic Medical Parasitology Workshop 2013 UPDATE – QUIZ PART 7 ORGANISM IDENTIFICATIONS,

ANSWER 3

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PVA stands for polyvinyl alcohol, a plastic powder/resin that is incorporated into the liquid fixative (Schaudinn’s or other fixatives) and serves as an adhesive to “glue” the stool material onto the slide. PVA itself has no preservation capability and is inert in terms of fixation, just like albumin.

TOTAL-FIX requires no adhesive (PVA or albumin).

Page 16: 2013 PARASITOLOGY WORKSHOP LYNNE S. GARCIA, MS, FAAM, CLS, BLM Diagnostic Medical Parasitology Workshop 2013 UPDATE – QUIZ PART 7 ORGANISM IDENTIFICATIONS,

ORGANISM # 9

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Blood Stain

Name genus, species and common name of disease

What’s a clue?

Page 17: 2013 PARASITOLOGY WORKSHOP LYNNE S. GARCIA, MS, FAAM, CLS, BLM Diagnostic Medical Parasitology Workshop 2013 UPDATE – QUIZ PART 7 ORGANISM IDENTIFICATIONS,

ORGANISM # 10

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Blood StainGenus, species

Page 18: 2013 PARASITOLOGY WORKSHOP LYNNE S. GARCIA, MS, FAAM, CLS, BLM Diagnostic Medical Parasitology Workshop 2013 UPDATE – QUIZ PART 7 ORGANISM IDENTIFICATIONS,

ORGANISM # 11

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Stool smear stained with Wheatley’s Trichrome stain.Genus, species

Page 19: 2013 PARASITOLOGY WORKSHOP LYNNE S. GARCIA, MS, FAAM, CLS, BLM Diagnostic Medical Parasitology Workshop 2013 UPDATE – QUIZ PART 7 ORGANISM IDENTIFICATIONS,

ORGANISM # 12

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Modified Trichrome StainGeneral term for organisms

Page 20: 2013 PARASITOLOGY WORKSHOP LYNNE S. GARCIA, MS, FAAM, CLS, BLM Diagnostic Medical Parasitology Workshop 2013 UPDATE – QUIZ PART 7 ORGANISM IDENTIFICATIONS,

QUESTION 4

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What are some of the immunoassay options available for stool protozoa?

How do they rank in terms of sensitivity and specificity?

Page 21: 2013 PARASITOLOGY WORKSHOP LYNNE S. GARCIA, MS, FAAM, CLS, BLM Diagnostic Medical Parasitology Workshop 2013 UPDATE – QUIZ PART 7 ORGANISM IDENTIFICATIONS,

RESPONSE 4

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All of the fecal immunoassay test formats (FA, EIA, Rapids) are comparable in terms of sensitivity and specificity. Selection of a particular method depends on work flow and the number of test requests and personal preference of that particular laboratory.

Page 22: 2013 PARASITOLOGY WORKSHOP LYNNE S. GARCIA, MS, FAAM, CLS, BLM Diagnostic Medical Parasitology Workshop 2013 UPDATE – QUIZ PART 7 ORGANISM IDENTIFICATIONS,

ORGANISM # 13

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Stool smear stained with Wheatley’s Trichrome stain.

Name genus, species, and stages

Page 23: 2013 PARASITOLOGY WORKSHOP LYNNE S. GARCIA, MS, FAAM, CLS, BLM Diagnostic Medical Parasitology Workshop 2013 UPDATE – QUIZ PART 7 ORGANISM IDENTIFICATIONS,

ORGANISM # 14

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Blood Stain; genus, species

Page 24: 2013 PARASITOLOGY WORKSHOP LYNNE S. GARCIA, MS, FAAM, CLS, BLM Diagnostic Medical Parasitology Workshop 2013 UPDATE – QUIZ PART 7 ORGANISM IDENTIFICATIONS,

QUESTION 5

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If Plasmodium falciparum parasites are in the capillaries, why not do a finger stick, rather than a venipuncture?

What is the best anticoagulant to use for blood specimens?

When should blood specimens be drawn for a suspect malaria diagnosis?

Page 25: 2013 PARASITOLOGY WORKSHOP LYNNE S. GARCIA, MS, FAAM, CLS, BLM Diagnostic Medical Parasitology Workshop 2013 UPDATE – QUIZ PART 7 ORGANISM IDENTIFICATIONS,

ANSWER 5

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The capillaries tend to be in the deep tissues (spleen, liver, bone marrow), so a finger stick sample is no more likely to be positive than a venipuncture blood.

EDTA is recommended as providing better organism morphology, particularly for Plasmodium spp.

Most patients we see in the US have never been exposed to malaria before, thus they have no antibody. These immunologically naïve patients may present with nonspecific symptoms that can mimic many other diseases. The rule of thumb is to draw immediately.

Page 26: 2013 PARASITOLOGY WORKSHOP LYNNE S. GARCIA, MS, FAAM, CLS, BLM Diagnostic Medical Parasitology Workshop 2013 UPDATE – QUIZ PART 7 ORGANISM IDENTIFICATIONS,

ORGANISM # 15

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Modified Acid-Fast Stain: What are the 3 structures shown in the slide (from largest to smallest)? The range is 10 µm to 2 µm.

Page 27: 2013 PARASITOLOGY WORKSHOP LYNNE S. GARCIA, MS, FAAM, CLS, BLM Diagnostic Medical Parasitology Workshop 2013 UPDATE – QUIZ PART 7 ORGANISM IDENTIFICATIONS,

ORGANISM # 16

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Iodine wet mount; seen on high dry power, X400What are morphologic characteristics that help you to identify this structure?

Page 28: 2013 PARASITOLOGY WORKSHOP LYNNE S. GARCIA, MS, FAAM, CLS, BLM Diagnostic Medical Parasitology Workshop 2013 UPDATE – QUIZ PART 7 ORGANISM IDENTIFICATIONS,

ORGANISM # 17

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India Ink stain: What and how do you count and how does this help to identify the structure?Name genus, species.

Page 29: 2013 PARASITOLOGY WORKSHOP LYNNE S. GARCIA, MS, FAAM, CLS, BLM Diagnostic Medical Parasitology Workshop 2013 UPDATE – QUIZ PART 7 ORGANISM IDENTIFICATIONS,

ORGANISM # 18

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Iodine wet mount: Seen on high dry power, X400. What are structures at each end called?Name genus, species

Page 30: 2013 PARASITOLOGY WORKSHOP LYNNE S. GARCIA, MS, FAAM, CLS, BLM Diagnostic Medical Parasitology Workshop 2013 UPDATE – QUIZ PART 7 ORGANISM IDENTIFICATIONS,

ORGANISM # 19

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Iodine wet mount: Identify genus and species – what is structure called in the circle?

O

Page 31: 2013 PARASITOLOGY WORKSHOP LYNNE S. GARCIA, MS, FAAM, CLS, BLM Diagnostic Medical Parasitology Workshop 2013 UPDATE – QUIZ PART 7 ORGANISM IDENTIFICATIONS,

ORGANISM # 20

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Iodine wet mount: What are the structures within the large object that help you to identify the genus and species?

Page 32: 2013 PARASITOLOGY WORKSHOP LYNNE S. GARCIA, MS, FAAM, CLS, BLM Diagnostic Medical Parasitology Workshop 2013 UPDATE – QUIZ PART 7 ORGANISM IDENTIFICATIONS,

QUESTION 6

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What are some of the key report comments associated with malaria reports?

What are the pros and cons of the thick and thin blood films?

Page 33: 2013 PARASITOLOGY WORKSHOP LYNNE S. GARCIA, MS, FAAM, CLS, BLM Diagnostic Medical Parasitology Workshop 2013 UPDATE – QUIZ PART 7 ORGANISM IDENTIFICATIONS,

PLASMODIUM SPP. Report Comments

¨ Plasmodium spp. seen: Unable to “rule out” Plasmodium falciparum.

¨ Plasmodium spp. not seen: One negative set of blood films will NOT “rule out” malaria; submit additional blood specimens every 4-6 hours.

¨ Plasmodium spp. seen, possible mixed infection: Unable to “rule out” Plasmodium falciparum.

¨ NOTE: It is mandatory that the parasitemia be calculated and reported for every initial and subsequent positive set of malarial films – the same method for determining parasitemia must be used for all blood film sets on that particular patient.

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Page 34: 2013 PARASITOLOGY WORKSHOP LYNNE S. GARCIA, MS, FAAM, CLS, BLM Diagnostic Medical Parasitology Workshop 2013 UPDATE – QUIZ PART 7 ORGANISM IDENTIFICATIONS,

THIN BLOOD FILMS

¨ Advantages– RBC morphology can be seen; RBCs preserved

– Compare size of infected RBCs to uninfected RBCs

– Much easier to identify to species level

– Easier to calculate parasitemia (%/100 RBCs)

¨ Disadvantages– Much lower sensitivity than thick blood film

– Infections with low parasitemia may be missed

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Page 35: 2013 PARASITOLOGY WORKSHOP LYNNE S. GARCIA, MS, FAAM, CLS, BLM Diagnostic Medical Parasitology Workshop 2013 UPDATE – QUIZ PART 7 ORGANISM IDENTIFICATIONS,

THICK BLOOD FILMS

¨ Advantages– Examining greater volume of blood– May be able to see malaria pigment within WBCs– May be able to see Schüffner's dots

¨ Disadvantages– Can’t compare sizes of infected and uninfected

RBCS– Organism distortion is difficult to recognize– Identification to species level is more difficult

GARCIA 35

Page 36: 2013 PARASITOLOGY WORKSHOP LYNNE S. GARCIA, MS, FAAM, CLS, BLM Diagnostic Medical Parasitology Workshop 2013 UPDATE – QUIZ PART 7 ORGANISM IDENTIFICATIONS,

ORGANISM # 21

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Cellophane tape: What are these structures and what is the collection method of choice?Name the genus and species

Page 37: 2013 PARASITOLOGY WORKSHOP LYNNE S. GARCIA, MS, FAAM, CLS, BLM Diagnostic Medical Parasitology Workshop 2013 UPDATE – QUIZ PART 7 ORGANISM IDENTIFICATIONS,

ORGANISM # 22

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Tissue biopsy, routine H/E stain: What is the cause of this infection? Why might bears, walrus, or pigs be involved?

Page 38: 2013 PARASITOLOGY WORKSHOP LYNNE S. GARCIA, MS, FAAM, CLS, BLM Diagnostic Medical Parasitology Workshop 2013 UPDATE – QUIZ PART 7 ORGANISM IDENTIFICATIONS,

THANKS – QUESTIONS?

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