Article information
Abstract
Amoebiasis is an infection caused by Entamoeba histolytica and is a potential health risk in countries in which health barriers are inappropriate. Since the discovery of Entamoeba dispar, the prevalence of amoebiasis has been modified.
ObjectiveThis study has standardized the PCR technique applied for the diagnosis of different species of the E. histolytica/E. dispar complex and has evaluated the prevalence of infection among patients attending private and public clinical laboratories in Salvador City, Bahia State, Brazil.
ResultsAnalysis of 52,704 stool samples by microscopic examination demonstrated that 1,788 (3.4%) were positive for the E. histolytica/E. dispar complex and infection occurred more often in samples originated from public clinical laboratories (5.0%) than those that came from private laboratories (3.2%). PCR performed in approximately 15% (262) E. histolytica/E. dispar complex positive samples, randomly chosen, amplified 227 samples (86.6%), all of them positive for E. dispar. The non-amplified 35 samples (13.4%) were also negative for E. histolytica-specific galactose adhesin. Moreover, to exclude a probable infection caused by E. hartmanni, morphometric analysis demonstrated that non-amplified samples had cyst sizes comparable to E. histolytica/ E. dispar (>10μm).
ConclusionThe absence of amplification of these samples indicates the presence of PCR inhibitors in the stool samples or the presence of DNA from Entamoeba species other than E. dispar, E. histolytica or E. hartmanni.
Keywords:
amebiasis
Entamoeba histolytica
diagnosis
prevalence
Full text is only aviable in PDF
References
[1.]
A. Caballero-Salcedo, M. Viveros-Rogel, B. Salvatierra, et al.
Seroepidemiology of amebiasis in Mexico.
Am J Trop Med Hyg, 50 (1994), pp. 412-419
[2.]
P. Kreidl, P. Imnadze, L. Baidoshvili, D. Greco.
Investigation of an outbreak of amoebiasis in Georgia.
Euro Surveil, 4 (1999), pp. 103-106
[3.]
S. Gatti, G. Swierczynski, F. Robinson, et al.
Amebic infections due to the Entamoeba histolytica-Entamoeba dispar complex: a study of the incidence in a remote rural area of Ecuador.
Am J Trop Med Hyg, 67 (2002), pp. 123-127
[4.]
M.L. Benetton, A.V. Gonçalves, M.E. Meneghini, et al.
Risk factors for infection by the Entamoeba histolytica/E. dispar complex: an epidemiological study conducted in outpatient clinics in the city of Manaus, Amazon region, Brazil.
Trans R Soc Trop Med Hyg, 99 (2005), pp. 532-540
[5.]
H.L. Santos, R.H. Peralta, H.W. de Macedo, et al.
Comparison of multiplex-PCR and antigen detection for differential diagnosis of Entamoeba histolytica.
Braz J Infect Dis, 11 (2007), pp. 365-370
[6.]
L. Mora, A. García, M. De Donato, H. Urdaneta.
Epidemiologic and molecular study of Entamoeba histolytica and Entamoeba dispar strains in patients with diarrhea in Cumana, Sucre state, Venezuela.
Invest Clin, 49 (2008), pp. 225-237
[7.]
World Health Organization.
Amoebiasis. Report on the WHO/Pan American Health Organization/UNESCO Expert Consultation, Mexico City. Geneva-WHO.
W Epidemiol Rec, 72 (1997), pp. 97-100
[8.]
N.J. Ashbolt.
Microbial contamination of drinking water and disease outcomes in developing regions.
Toxicology, 198 (2004), pp. 229-238
[9.]
B. Chávez, A. Martínez-Palomo, M. De La Torre.
Ultramicroscopic structure of the cyst wall of Entamoeba invadens, E. histolytica and E. coli.
Arch Invest Med, (1978), pp. 113-116
[10.]
L.S. Ritchie.
An ether sedimentation technique for routine stool examinations.
Bull U S Army Med Dep, 8 (1948), pp. 326
[11.]
R. Acuna-Soto, J. Samuelson, P. De Girolami, et al.
Application of the polymerase chain reaction to the epidemiology of pathogenic and nonpathogenic Entamoeba histolytica.
Am J Trop Med Hyg, 48 (1993), pp. 58-70
[12.]
A. Evangelopoulos, G. Spanakos, E. Patsoula, et al.
A nested, multiplex, PCR assay for the simultaneous detection and differentiation of Entamoeba histolytica and Entamoeba dispar in faeces.
Ann Trop Med Parasitol, 94 (2000), pp. 233-240
[13.]
J.J. Verweij, J. Blotkamp, E.A. Brienen, et al.
Differentiation of Entamoeba histolytica and Entamoeba dispar cysts using polymerase chain reaction on DNA isolated from faeces with spin columns.
Eur J Clin Microbiol Infect Dis, 19 (2000), pp. 358-361
[14.]
S. Zaman, J. Khoo, S.W. Ng, et al.
Direct amplification of Entamoeba histolytica DNA from amoebic liver abscess pus using polymerase chain reaction.
Parasitol Res, 86 (2000), pp. 724-728
[15.]
E.F. Silva, M.A. Gomes, et al.
Amebíase: Entamoeba histolytica/Entamoeba dispar.
Parasitologia Humana,
[16.]
R. Araújo, C.N.C. Bichara, L.C.L. Chaves, et al.
Doenças infecciosas e parasitárias – enfoque Amazônico.
CEJUP. Universidade Federal do Pará,
[17.]
W.A. Hoffman, J.A. Pons, J.L. Janer.
The sedimentation concentration method in schistosomiasis mansoni.
Puerto Rico J Publ Health Trop Med, 9 (1934), pp. 283-298
[18.]
Brazilian Institute of Geography and Statistics. Contagem da População, 2007. http://www.ibge.gov.br/home/estatistica/populacao/contagem2007/contagem_final/tabela1_1_16.pdf.
[19.]
F.L.N. Santos, A.N. Hiltner, F.G. Araújo, et al.
Prevalência e perfil epidemiológico das enteroparasitoses em Salvador (BA).
40° Congresso Brasileiro de Patologia Clínica e Medicina Laboratorial, Curitiba-PR, (2006),
[20.]
L. Rey.
Parasitologia: parasitos e doenças parasitárias do homem nos trópicos ocidentais..
Guanabara-Koogan Press, (2008),
[21.]
H. Troll, H. Marti, N. Weiss.
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 (1997), pp. 1701-1705
[22.]
B. Leiva, M. Lebbad, J. Winiecka-Krusnell, et al.
Overdiagnosis of Entamoeba histolytica and Entamoeba dispar in Nicaragua: a microscopic, triage parasite panel and PCR study.
Arch Med Res, 37 (2006), pp. 529-534
[23.]
P.R. Valle, M.B. Souza, E.M. Pires, et al.
Arbitrarily primed PCR fingerprint of RNA and DNA in Entamoeba histolytica.
Rev Inst Med Trop São Paulo, 42 (2000), pp. 249-253
[24.]
A. Evangelopoulos, N. Legakis, N. Vakalis.
Microscopy, PCR and ELISA applied to the epidemiology of amoebiasis in Greece.
Parasitol Int, 50 (2001), pp. 185-189
[25.]
S. Roy, M. Kabir, D. Mondal, et al.
Real-time-PCR assay for diagnosis of Entamoeba histolytica infection.
J Clin Microbiol, 43 (2005), pp. 2168-2172
[26.]
E.N. Vianna, J.O. Costa, C.K. Santos, et al.
An alternative method for DNA extraction and PCR identification of Entamoeba histolytica and E. dispar in fecal samples.
Parasitology, 136 (2009), pp. 765-769
[27.]
S. Zindrou, E. Orozco, E. Linder, et al.
Specific detection of Entamoeba histolytica DNA by hemolysin gene targeted PCR.
Acta Trop, 78 (2001), pp. 117-125
[28.]
A. Calderaro, C. Gorrini, S. Bommezzadri, et al.
Entamoeba histolytica and Entamoeba dispar: comparison of two PCR assays for diagnosis in a non-endemic setting.
Trans R Soc Trop Med Hyg, 100 (2006), pp. 450-457
[29.]
M. Tanyuksel, W.A. Petri Jr.
Laboratory diagnosis of amoebiasis.
Clin Microbiol Rev, 16 (2003), pp. 713-729
[30.]
R. Deuter, S. Pietsch, S. Hertel, O. Müller.
A method for preparation of fecal DNA suitable for PCR.
Nucleic Acids Res, 23 (1995), pp. 3800-3801
[31.]
P.G. Lantz, F. Tjerneld, B. Hahn-Hägerdal, P. Radström.
Use of aqueous two-phase systems in sample preparation for polymerase chain reaction-based detection of microorganisms.
J Chromatogr B Biomed Appl, 680 (1996), pp. 165-170
[32.]
N. Vandenberg, R.A. van Oorschot.
Extraction of human nuclear DNA from feces samples using the QIAamp DNA Stool Mini Kit.
J Forensic Sci, 47 (2002), pp. 993-995
[33.]
E. Campos-Góngora, F. Ebert, U. Willhoeft, et al.
Characterization of chitin synthases from Entamoeba.
Protist, 155 (2004), pp. 323-330
[34.]
L. Monteiro, D. Bonnemaison, A. Vekris, et al.
Complex polysaccharides as PCR inhibitors in feces: Helicobacter pylori model.
J Clin Microbiol, 35 (1997), pp. 995-998
[35.]
I.G. Wilson.
Inhibition and facilitation of nucleic acid amplification.
Appl Environ Microbiol, 63 (1997), pp. 3741-3751
[36.]
A.S. Cunha, E.F. Silva, T.C.A. Ferrari, et al.
Amebíase em tópicos e gastroenterologia.
Amebiases,
[37.]
J. Oliveira-Costa, J.A. Resende, M.A. Gomes.
Prevalência de Entamoeba histolytica e Entamoeba dispar em amostras fecais de um laboratório privado na grande Belo Horizonte.
Semana de Iniciação Científica da Universidade Federal de Minas Gerais, Belo Horizonte, MG, (2006),
[38.]
A. Dourado, A. Maciel, S. Aca Ida.
Occurrence of Entamoeba histolytica/Entamoeba dispar in ambulatory patients of Recife, PE.
Rev Soc Bras Med Trop, 39 (2006), pp. 388-389
Copyright © 2011. Elsevier Editora Ltda.. All rights reserved
