Application of Real-Time Polymerase Chain Reaction in Detection of<i>Entamoeba histolytica</i>in Pus Aspirates of Liver Abscess Patients

Othman, Nurulhasanah; Mohamed, Zeehaida; Verweij, Jaco J.; Lim, Boon Huat; Olivos-García, Alfonso; Chen, Yeng; Noordin, Rahmah

doi:10.1089/fpd.2009.0427

Cited by 33 publications

(20 citation statements)

References 18 publications

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“…The procedures of collecting and handling the serum samples were approved by USM Human Ethical Committee (reference number USMKK/PPP/ JEPem[213.3(10)]). Human serum samples included in this study were divided into four groups: (i) group A, human ALA serum samples (n ϭ 24) with consistent clinical symptoms (i.e., fever, abdominal/right hepatic chest pain, hepatomegaly, and jaundice) and radiological image and IHA-positive results; (ii) group B, human ALA serum samples (n ϭ 14) from patients whose abscesses were positive by real-time PCR for E. histolytica DNA and negative by bacterial culture (the primers and probe sequences were as reported previously [9]); (iii) group C, healthy blood donor serum samples which were negative by IHA (n ϭ 30); (iv) group D, serum samples from patients with other infections (n ϭ 33), i.e., salmonellosis (n ϭ 5), shigellosis (n ϭ 1), Escherichia coli septicemia (n ϭ 2), Staphylococcus sp. septicemia (n ϭ 2), Helicobacter pylori (n ϭ 6), pyogenic liver abscess (n ϭ 4), Stenotrophomonas maltophilia septicemia (n ϭ 1), enteropathogenic E. coli (n ϭ 1), Ascaris lumbricoides (n ϭ 1), Klebsiella pneumoniae (n ϭ 1), and toxoplasmosis (n ϭ 9).…”

Section: Methodsmentioning

confidence: 99%

Analysis of Entamoeba histolytica Excretory-Secretory Antigen and Identification of a New Potential Diagnostic Marker

Kin

Tan

Othman

et al. 2011

Clin Vaccine Immunol

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Serodiagnosis of amoebiasis remains the preferred method for diagnosis of amoebic liver abscess (ALA).However, the commercially available kits are problematic in areas of endemicity due to the persistently high background antibody titers. Human serum samples (n ‫؍‬ 38) from patients with ALA who live in areas of endemicity were collected from Hospital Universiti Sains Malaysia during the period of 2008 to 2010. Western blots using excretory-secretory antigen (ESA) collected from axenically grown Entamoeba histolytica were probed with the above serum samples. Seven antigenic proteins of ESA with various reactivities were identified, i.e., 152 kDa, 131 kDa, 123 kDa, 110 kDa, 100 kDa, 82 kDa, and 76 kDa. However, only the 152-kDa and 110-kDa proteins showed sensitivities above 80% in the Western blot analysis. All the antigenic proteins showed undetectable cross-reactivity when probed with healthy human serum samples (n ‫؍‬ 30) and serum samples from other infections (n ‫؍‬ 33). From the matrix-assisted laser desorption ionization-two-stage time of flight (MALDI-TOF/TOF) analysis, the proteins were identified as heavy subunits of E. histolytica lectin and E. histolytica pyruvate phosphate dikinase, respectively. Use of the E. histolytica lectin for diagnosis of ALA has been well reported by researchers and is being used in commercialized kits. However, this is the first report on the potential use of pyruvate phosphate dikinase for diagnosis of ALA; thus, this molecule merits further evaluation on its diagnostic value using a larger panel of serum samples.

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Section: Methodsmentioning

confidence: 99%

Analysis of Entamoeba histolytica Excretory-Secretory Antigen and Identification of a New Potential Diagnostic Marker

Kin

Tan

Othman

et al. 2011

Clin Vaccine Immunol

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“…PCR technology also facilitates efforts toward identification of E. histolytica in samples other than stool, e.g., aspirates from liver abscesses, cerebrospinal fluid (CSF), and urine (51,(68)(69)(70). The wider availability of real-time PCR platforms has paved the way for the routine use of PCRs for this and other parasite targets as first-line diagnostic methods.…”

Section: Entamoebamentioning

confidence: 99%

Molecular Testing for Clinical Diagnosis and Epidemiological Investigations of Intestinal Parasitic Infections

2014

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SUMMARY Over the past few decades, nucleic acid-based methods have been developed for the diagnosis of intestinal parasitic infections. Advantages of nucleic acid-based methods are numerous; typically, these include increased sensitivity and specificity and simpler standardization of diagnostic procedures. DNA samples can also be stored and used for genetic characterization and molecular typing, providing a valuable tool for surveys and surveillance studies. A variety of technologies have been applied, and some specific and general pitfalls and limitations have been identified. This review provides an overview of the multitude of methods that have been reported for the detection of intestinal parasites and offers some guidance in applying these methods in the clinical laboratory and in epidemiological studies.

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“…They were nevertheless, found to be inadequate in highly endemic regions and new detection methodologies such as antigen detection ELISA and polymerase chain reaction (PCR) have since been developed Othman et al 2010;Ackers 2002;Tanyuksel and Petri 2003). Both techniques are widely used nowadays for detecting and identifying E. histolytica in clinical samples.…”

Section: Introductionmentioning

confidence: 99%

Differential detection of Entamoeba histolytica, Entamoeba dispar and Entamoeba moshkovskii in fecal samples by nested PCR in the United Arab Emirates (UAE)

ElBakri¹,

Samie²,

Ezzedine³

et al. 2013

Acta Parasitologica

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Amoebiasis is one of the most important infectious diseases afflicting mainly tropical and subtropical countries. This study was carried out in the Sharjah Emirate, UAE in order to accurately detect and differentiate Entamoeba histolytica, Entamoeba dispar and E. moshkovskii in fecal samples collected from the Sharjah municipality public health clinic by ELISA and nested polymerase chain reaction (PCR). One hundred and twenty specimens were examined and the PCR was positive for E. histolytica, E. dispar and E. moshkovskii (collectively referred to as Entamoeba complex) in 19.2% (23 out of 120). Of those, 10% (12/120) were mono -infection with E. histolytica; 2.5% (3/120) with E. dispar; and 2.5% (3/120) E. moshkovskii. The nested PCR also detected mixed infections by both E. histolytica and E. dispar in 3.3% (4/120) and E. dispar and E. moshkovskii in 0.8% (1/120). The TechLab ELISA kit failed to detect E. histolytica in any of the E. histolytica PCR positive samples. Overall, the percentage of E. histolytica including those found in mixed infections was 13.3% (16/120). Compared to nested PCR, microscopy was found to have an overall sensitivity of 52.2% and a specificity of 75.2% for detection of Entamoeba complex. The present study indicates that E. histolytica is present in the UAE with an average incidence rate of 13.3%. However, larger studies need to be conducted in order to confirm these findings. We propose the use of PCR in both the routine diagnosis of amoebiasis and epidemiological survey in the UAE.

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Application of Real-Time Polymerase Chain Reaction in Detection ofEntamoeba histolyticain Pus Aspirates of Liver Abscess Patients

Cited by 33 publications

References 18 publications

Analysis of Entamoeba histolytica Excretory-Secretory Antigen and Identification of a New Potential Diagnostic Marker

Analysis of Entamoeba histolytica Excretory-Secretory Antigen and Identification of a New Potential Diagnostic Marker

Molecular Testing for Clinical Diagnosis and Epidemiological Investigations of Intestinal Parasitic Infections

Differential detection of Entamoeba histolytica, Entamoeba dispar and Entamoeba moshkovskii in fecal samples by nested PCR in the United Arab Emirates (UAE)

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