Amoebic liver abscess other diagnostic studies: Difference between revisions

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* Presence of RBCs in trophozoites is diagnostic of Entamoeba histolytica
* Presence of RBCs in trophozoites is diagnostic of Entamoeba histolytica
* The specificity of the test is low as the trophozoites containing RBCs are not present in all cases and E dispar may also contain RBCs in trophozoites<ref name="pmid8163695">{{cite journal| author=González-Ruiz A, Haque R, Aguirre A, Castañón G, Hall A, Guhl F et al.| title=Value of microscopy in the diagnosis of dysentery associated with invasive Entamoeba histolytica. | journal=J Clin Pathol | year= 1994 | volume= 47 | issue= 3 | pages= 236-9 | pmid=8163695 | doi= | pmc=501902 | url=https://www.ncbi.nlm.nih.gov/entrez/eutils/elink.fcgi?dbfrom=pubmed&tool=sumsearch.org/cite&retmode=ref&cmd=prlinks&id=8163695  }} </ref><ref name="pmid2894495">{{cite journal| author=Strachan WD, Chiodini PL, Spice WM, Moody AH, Ackers JP| title=Immunological differentiation of pathogenic and non-pathogenic isolates of Entamoeba histolytica. | journal=Lancet | year= 1988 | volume= 1 | issue= 8585 | pages= 561-3 | pmid=2894495 | doi= | pmc= | url=https://www.ncbi.nlm.nih.gov/entrez/eutils/elink.fcgi?dbfrom=pubmed&tool=sumsearch.org/cite&retmode=ref&cmd=prlinks&id=2894495  }} </ref>
* The specificity of the test is low as the trophozoites containing RBCs are not present in all cases and E dispar may also contain RBCs in trophozoites<ref name="pmid8163695">{{cite journal| author=González-Ruiz A, Haque R, Aguirre A, Castañón G, Hall A, Guhl F et al.| title=Value of microscopy in the diagnosis of dysentery associated with invasive Entamoeba histolytica. | journal=J Clin Pathol | year= 1994 | volume= 47 | issue= 3 | pages= 236-9 | pmid=8163695 | doi= | pmc=501902 | url=https://www.ncbi.nlm.nih.gov/entrez/eutils/elink.fcgi?dbfrom=pubmed&tool=sumsearch.org/cite&retmode=ref&cmd=prlinks&id=8163695  }} </ref><ref name="pmid2894495">{{cite journal| author=Strachan WD, Chiodini PL, Spice WM, Moody AH, Ackers JP| title=Immunological differentiation of pathogenic and non-pathogenic isolates of Entamoeba histolytica. | journal=Lancet | year= 1988 | volume= 1 | issue= 8585 | pages= 561-3 | pmid=2894495 | doi= | pmc= | url=https://www.ncbi.nlm.nih.gov/entrez/eutils/elink.fcgi?dbfrom=pubmed&tool=sumsearch.org/cite&retmode=ref&cmd=prlinks&id=2894495  }} </ref>
*Freezing a fresh fecal specimen at -20°C before extraction of DNA will not effect the sensitivity of the molecular assays.<ref name="pmid17229864">{{cite journal| author=Fotedar R, Stark D, Beebe N, Marriott D, Ellis J, Harkness J| title=PCR detection of Entamoeba histolytica, Entamoeba dispar, and Entamoeba moshkovskii in stool samples from Sydney, Australia. | journal=J Clin Microbiol | year= 2007 | volume= 45 | issue= 3 | pages= 1035-7 | pmid=17229864 | doi=10.1128/JCM.02144-06 | pmc=1829108 | url=https://www.ncbi.nlm.nih.gov/entrez/eutils/elink.fcgi?dbfrom=pubmed&tool=sumsearch.org/cite&retmode=ref&cmd=prlinks&id=17229864  }} </ref><ref name="pmid16126570">{{cite journal| author=Lebbad M, Svärd SG| title=PCR differentiation of Entamoeba histolytica and Entamoeba dispar from patients with amoeba infection initially diagnosed by microscopy. | journal=Scand J Infect Dis | year= 2005 | volume= 37 | issue= 9 | pages= 680-5 | pmid=16126570 | doi=10.1080/00365540510037812 | pmc= | url=https://www.ncbi.nlm.nih.gov/entrez/eutils/elink.fcgi?dbfrom=pubmed&tool=sumsearch.org/cite&retmode=ref&cmd=prlinks&id=16126570  }} </ref><ref name="pmid11463120">{{cite journal| author=Núñez YO, Fernández MA, Torres-Núñez D, Silva JA, Montano I, Maestre JL et al.| title=Multiplex polymerase chain reaction amplification and differentiation of Entamoeba histolytica and Entamoeba dispar DNA from stool samples. | journal=Am J Trop Med Hyg | year= 2001 | volume= 64 | issue= 5-6 | pages= 293-7 | pmid=11463120 | doi= | pmc= | url=https://www.ncbi.nlm.nih.gov/entrez/eutils/elink.fcgi?dbfrom=pubmed&tool=sumsearch.org/cite&retmode=ref&cmd=prlinks&id=11463120  }} </ref>
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|'''Wet (Saline) preparation'''
|'''Wet (Saline) preparation'''

Revision as of 17:55, 10 February 2017

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Editor-In-Chief: C. Michael Gibson, M.S., M.D. [1];Associate Editor(s)-in-Chief: Yamuna Kondapally, M.B.B.S[2]

Overview

Other Diagnostic Tests

Needle aspiration

  • Needle aspiration is used to differentiate between amoebic and pyogenic liver abscess
  • The ultrasound and CT are used to guide percutaneous aspiration and drainage.
  • The aspirated fluid is odorless unless secondarily infected
  • The pathognomonic of the aspirate is the reddish-brown anchovy paste appearance, which indicates that the abscess has been present for weeks.
  • The trophozoites are present only in the wall of the abscess. Rest of abscess is composed of lysed leukocytes.
Laboratory Method Findings
Microscopy
  • Microscopic techniques include 1. Wet preparation 2. Concentration 3. Permanently stained smears[1]
  • Less reliable method than culture or antigen detection test[2][3]
  • Minimum three stool samples are examined for ova and parasites within 10days of sample collection as organisms are excreted intermittently[4]
  • Presence of RBCs in trophozoites is diagnostic of Entamoeba histolytica
  • The specificity of the test is low as the trophozoites containing RBCs are not present in all cases and E dispar may also contain RBCs in trophozoites[5][6]
  • Freezing a fresh fecal specimen at -20°C before extraction of DNA will not effect the sensitivity of the molecular assays.[7][8][9]
Wet (Saline) preparation
  • Very insensitive method (<10%) Procedure
  • The sample is a fresh specimen that should be examined within 1 hr of collection
  • The test is positive when RBCs in trophozoites are detected
  • It is not used in patients without acute dysentery as trophozoites will not contain RBCs
 Concentration Technique
  • It is helpful in detecting cysts in the stool sample in asymptomatic carriers
 Permanently stained smears
  • This is an important method for identification and recovery of Entamoeba species
Culture Methods
  • The specimens for the culture of E histolytica include fecal specimens, rectal biopsy specimens or liver abscess aspirates.
  • There are two culture techniques. They are xenic and axenic systems.[10]
  • Culture of E histolytica is less sensitive than microscopy as a detection method.
  • Parasite cultures are expensive, labor intensive and difficult to maintain in the diagnostic laboratory.

Disadvantage

  • Culture is not recommended as the routine diagnostic test due to overgrowth of other organisms like bacteria and fungi in the culture media.
Isoenzyme analysis
  • Isoenzyme analysis is useful in the differentiation of Entamoeba species.[11][12][13]
  • There are three isoenzymes (zymodyme) for E histolytica and one for E dispar.
  • This techniques helps in differentiation of E histolytica and E dispar and is considered gold standard for diagnosing amoebic infection prior to development of newer DNA-based techniques.

Disadvantages

  • Difficult to perform test, time-consuming procedure, and not always successful.
  • The isoenzyme analysis is negative for many microscopy positive stool samples[5][14][15][6]
Antibody Detection Tests
  • Antibody detection tests are useful in the case of amoebic liver abscess as patients do not have detectable parasites in feces.
  • The sensitivity of the test is 100% in patients with amoebic liver abscess. The techniques include enzyme-linked immunosorbent assay (ELISA), indirect hemagglutination (IHA), latex agglutination, immunoelectrophoresis, counterimmunoelectrophoresis (CIE), the amebic gel diffusion test, immunodiffusion, complement fixation, and indirect immunofluorescence assay (IFA).

ELISA

  • Useful in the diagnosis of asymptomatic and symptomatic amoebiasis after fecal examination.
  • Useful in amoebic liver abscess and in the evaluation of intestinal and extraintestinal infections where organisms cannot be detected in feces but amebiasis is suspected.[16]
  • Easy to perform in the clinical laboratory.
  • The presence of IgM antibodies indicates current infection and IgG antibodies persists for years after E histolytica infection.[17]
  • Presence of antilectin antibodies are frequently used for the diagnosis of patients with amoebic liver abscess.[18]
  • The sensitivity is 95%. It is a useful test for the diagnostic clinical laboratory as this test does not have cross reaction with other non-Entamoeba histolytica species.[19][20][21][22][2]
Antigen Detection Tests
  • The antigen detection tests use monoclonal antibodies against the Gal/GalNAc-specific lectin of E. histolytica or against serine-rich antigen of E. histolytica.
  • This test is useful in differentiation of disease causing E histolytica as there are antigenic differences in the lectins of E histolytica and E dispar.
  • The test has good sensitivity and specificity for the detection of E histolytica antigen in stool specimens of patients with amoebic colitis and asymptomatic intestinal infection.[23][14][15]
  • Antigen detection using ELISA is is both rapid and technically simple to perform. Hence used in developing countries where amoebiasis is most prevalent.

Disadvantage

  • Antigens detected are denatured by fixation of the stool sample. hence the test is limited to frozen or fresh samples.
Immunochromatographic Assays
  • The triage parasite panel (TPP) is used for antigen detection of antigens specific for E. histolytica/E. dispar, Giardia lamblia, and Cryptosporidium parvum simultaneously.[24][25]
  • The antigens specific for these organisms are used by using specific antibodies and immobilized on a membrane.
  • This test is highly sensitive and specific.
  • This test can be performed within 15 min with fresh or frozen, unfixed human fecal specimens.

Disadvantage

  • This test does not differentiate between E histolytica and E dispar. Hence not a method of choice for the diagnostic laboratory.
  • Stool samples are transported to the laboratory as soon as possible as the test is performed on the fresh or fresh-frozen unpreserved stool samples.[26][27]

DNA-Based Diagnostic Tests

  • These tests are limited to developed countries in research and clinical laboratories.
  • Fecal specimens for DNA analysis may be preserved by refrigeration or in a formalin, SAF or PVA fixative. Formalin preserves cysts, and PVA and SAF preserves trophozoites and cysts in wet mounts.[28][29]
  • the sample medium which is used to transport amoebic DNA is ethanol and the reagent used for the preservation of fecal samples is 10% buffered formalin solution.[30]
Laboratory Methods Findings
Manual Methods
Automated Methods
Conventional PCR
Real-Time PCR
Microarray Development
Typing Methods

References

  1. ↑ Huston CD, Haque R, Petri WA (1999). "Molecular-based diagnosis of Entamoeba histolytica infection". Expert Rev Mol Med. 1999: 1–11. doi:doi:10.1017/S1462399499000599 Check |doi= value (help). PMID 14987356.
  2. ↑ 2.0 2.1 Tanyuksel M, Petri WA (2003). "Laboratory diagnosis of amebiasis". Clin Microbiol Rev. 16 (4): 713–29. PMC 207118. PMID 14557296.
  3. ↑ Krogstad DJ, Spencer HC, Healy GR, Gleason NN, Sexton DJ, Herron CA (1978). "Amebiasis: epidemiologic studies in the United States, 1971-1974". Ann Intern Med. 88 (1): 89–97. PMID 619763.
  4. ↑ Li E, Stanley SL (1996). "Protozoa. Amebiasis". Gastroenterol Clin North Am. 25 (3): 471–92. PMID 8863036.
  5. ↑ 5.0 5.1 González-Ruiz A, Haque R, Aguirre A, Castañón G, Hall A, Guhl F; et al. (1994). "Value of microscopy in the diagnosis of dysentery associated with invasive Entamoeba histolytica". J Clin Pathol. 47 (3): 236–9. PMC 501902. PMID 8163695.
  6. ↑ 6.0 6.1 Strachan WD, Chiodini PL, Spice WM, Moody AH, Ackers JP (1988). "Immunological differentiation of pathogenic and non-pathogenic isolates of Entamoeba histolytica". Lancet. 1 (8585): 561–3. PMID 2894495.
  7. ↑ Fotedar R, Stark D, Beebe N, Marriott D, Ellis J, Harkness J (2007). "PCR detection of Entamoeba histolytica, Entamoeba dispar, and Entamoeba moshkovskii in stool samples from Sydney, Australia". J Clin Microbiol. 45 (3): 1035–7. doi:10.1128/JCM.02144-06. PMC 1829108. PMID 17229864.
  8. ↑ Lebbad M, Svärd SG (2005). "PCR differentiation of Entamoeba histolytica and Entamoeba dispar from patients with amoeba infection initially diagnosed by microscopy". Scand J Infect Dis. 37 (9): 680–5. doi:10.1080/00365540510037812. PMID 16126570.
  9. ↑ Núñez YO, Fernández MA, Torres-Núñez D, Silva JA, Montano I, Maestre JL; et al. (2001). "Multiplex polymerase chain reaction amplification and differentiation of Entamoeba histolytica and Entamoeba dispar DNA from stool samples". Am J Trop Med Hyg. 64 (5–6): 293–7. PMID 11463120.
  10. ↑ Clark CG, Diamond LS (2002). "Methods for cultivation of luminal parasitic protists of clinical importance". Clin Microbiol Rev. 15 (3): 329–41. PMC 118080. PMID 12097242.
  11. ↑ Sargeaunt PG, Jackson TF, Wiffen S, Bhojnani R, Williams JE, Felmingham D; et al. (1987). "The reliability of Entamoeba histolytica zymodemes in clinical laboratory diagnosis". Arch Invest Med (Mex). 18 (2): 69–75. PMID 2889435.
  12. ↑ Blanc D, Sargeaunt PG (1991). "Entamoeba histolytica zymodemes: exhibition of gamma and delta bands only of glucose phosphate isomerase and phosphoglucomutase may be influenced by starch content in the medium". Exp Parasitol. 72 (1): 87–90. PMID 1825198.
  13. ↑ Jackson TF, Suparsad S (1997). "Zymodeme stability of Entamoeba histolytica and E. dispar". Arch Med Res. 28 Spec No: 304–5. PMID 9033111.
  14. ↑ 14.0 14.1 Haque R, Faruque AS, Hahn P, Lyerly DM, Petri WA (1997). "Entamoeba histolytica and Entamoeba dispar infection in children in Bangladesh". J Infect Dis. 175 (3): 734–6. PMID 9041357.
  15. ↑ 15.0 15.1 Haque R, Neville LM, Hahn P, Petri WA (1995). "Rapid diagnosis of Entamoeba infection by using Entamoeba and Entamoeba histolytica stool antigen detection kits". J Clin Microbiol. 33 (10): 2558–61. PMC 228528. PMID 8567882.
  16. ↑ Rosenblatt JE, Sloan LM, Bestrom JE (1995). "Evaluation of an enzyme-linked immunoassay for the detection in serum of antibodies to Entamoeba histolytica". Diagn Microbiol Infect Dis. 22 (3): 275–8. PMID 8565416.
  17. ↑ Abd-Alla MD, Jackson TG, Ravdin JI (1998). "Serum IgM antibody response to the galactose-inhibitable adherence lectin of Entameoba histolytica". Am J Trop Med Hyg. 59 (3): 431–4. PMID 9749639.
  18. ↑ Ravdin JI, Jackson TF, Petri WA, Murphy CF, Ungar BL, Gathiram V; et al. (1990). "Association of serum antibodies to adherence lectin with invasive amebiasis and asymptomatic infection with pathogenic Entamoeba histolytica". J Infect Dis. 162 (3): 768–72. PMID 2388003.
  19. ↑ Braga LL, Mendonca Y, Paiva CA, Sales A, Cavalcante AL, Mann BJ (1998). "Seropositivity for and intestinal colonization with Entamoeba histolytica and entamoeba dispar in individuals in northeastern Brazil". J Clin Microbiol. 36 (10): 3044–5. PMC 105108. PMID 9738064.
  20. ↑ Goncalves ML, da Silva VL, de Andrade CM, Reinhard K, da Rocha GC, Le Bailly M; et al. (2004). "Amoebiasis distribution in the past: first steps using an immunoassay technique". Trans R Soc Trop Med Hyg. 98 (2): 88–91. PMID 14964807.
  21. ↑ Grundy MS (1982). "Preliminary observations using a multi-layer ELISA method for the detection of Entamoeba histolytica trophozoite antigens in stool samples". Trans R Soc Trop Med Hyg. 76 (3): 396–400. PMID 6287685.
  22. ↑ Shamsuzzaman SM, Haque R, Hasin SK, Hashiguchi Y (2000). "Evaluation of indirect fluorescent antibody test and enzyme-linked immunosorbent assay for diagnosis of hepatic amebiasis in Bangladesh". J Parasitol. 86 (3): 611–5. doi:10.1645/0022-3395(2000)086[0611:EOIFAT]2.0.CO;2. PMID 10864262.
  23. ↑ Haque R, Ali IK, Akther S, Petri WA (1998). "Comparison of PCR, isoenzyme analysis, and antigen detection for diagnosis of Entamoeba histolytica infection". J Clin Microbiol. 36 (2): 449–52. PMC 104557. PMID 9466756.
  24. ↑ Leiva B, Lebbad M, Winiecka-Krusnell J, Altamirano I, Tellez A, Linder E (2006). "Overdiagnosis of Entamoeba histolytica and Entamoeba dispar in Nicaragua: a microscopic, triage parasite panel and PCR study". Arch Med Res. 37 (4): 529–34. doi:10.1016/j.arcmed.2005.10.009. PMID 16624654.
  25. ↑ Pillai DR, Kain KC (1999). "Immunochromatographic strip-based detection of Entamoeba histolytica-E. dispar and Giardia lamblia coproantigen". J Clin Microbiol. 37 (9): 3017–9. PMC 85440. PMID 10449494.
  26. ↑ Garcia LS, Shimizu RY, Bernard CN (2000). "Detection of Giardia lamblia, Entamoeba histolytica/Entamoeba dispar, and Cryptosporidium parvum antigens in human fecal specimens using the triage parasite panel enzyme immunoassay". J Clin Microbiol. 38 (9): 3337–40. PMC 87383. PMID 10970380.
  27. ↑ Sharp SE, Suarez CA, Duran Y, Poppiti RJ (2001). "Evaluation of the Triage Micro Parasite Panel for detection of Giardia lamblia, Entamoeba histolytica/Entamoeba dispar, and Cryptosporidium parvum in patient stool specimens". J Clin Microbiol. 39 (1): 332–4. doi:10.1128/JCM.39.1.332-334.2001. PMC 87724. PMID 11136793.
  28. ↑ Ramos F, Zurabian R, Morán P, Ramiro M, Gómez A, Clark CG; et al. (1999). "The effect of formalin fixation on the polymerase chain reaction characterization of Entamoeba histolytica". Trans R Soc Trop Med Hyg. 93 (3): 335–6. PMID 10492774.
  29. ↑ Troll H, Marti H, Weiss N (1997). "Simple differential detection of Entamoeba histolytica and Entamoeba dispar in fresh stool specimens by sodium acetate-acetic acid-formalin concentration and PCR". J Clin Microbiol. 35 (7): 1701–5. PMC 229825. PMID 9196177.
  30. ↑ https://www.cdc.gov/dpdx/reference.html Accessed on February 10, 2017
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