Detection and Identification of
            <i>Entamoeba</i>
            Species in Stool Samples by a Reverse Line Hybridization Assay

Verweij, Jaco J.; Laeijendecker, Daphne; Brienen, Eric A. T.; Lieshout, Lisette van; Polderman, A.M.

doi:10.1128/jcm.41.11.5041-5045.2003

Cited by 48 publications

(26 citation statements)

References 12 publications

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“…This simple diagnostic PCR technique does not require extra steps, as is the case with nested PCR (2), PCRrestricted fragment length polymorphism (13), and PCR with reverse line blot hybridization (14).…”

Section: Discussionmentioning

confidence: 99%

“…The sensitivity of these methods for detecting E. moshkovskii has been shown to be higher than that of microscopy. However, these methods are still relatively time-consuming and require extra and complex procedures, such as nested PCR, restriction endonuclease assays, or hybridization, to achieve their higher sensitivity (2,3,14).…”

mentioning

confidence: 99%

“…PCR coupled with additional methods, such as PCR with restriction fragment length polymorphism analysis or riboprinting (3) and PCR with reverse line blot hybridization (14), has also been used to differentiate E. moshkovskii from other species. The sensitivity of these methods for detecting E. moshkovskii has been shown to be higher than that of microscopy.…”

mentioning

confidence: 99%

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Differential Detection of Entamoeba histolytica , Entamoeba dispar , and Entamoeba moshkovskii by a Single-Round PCR Assay

Hamzah¹,

et al. 2006

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A single-round PCR assay was developed for detection and differential diagnosis of the three Entamoeba species found in humans, Entamoeba moshkovskii, Entamoeba histolytica, and Entamoeba dispar, that are morphologically identical as both cysts and trophozoites. A conserved forward primer was derived from the middle of the small-subunit rRNA gene, and reverse primers were designed from signature sequences specific to each of these three Entamoeba species. PCR generates a 166-bp product with E. histolytica DNA, a 752-bp product with E. dispar DNA, and a 580-bp product with E. moshkovskii DNA. Thirty clinical specimens were examined, and the species present were successfully detected and differentiated using this assay. It was possible to detect as little as 10 pg of E. moshkovskii and E. histolytica DNA, while for E. dispar the sensitivity was about 20 pg of DNA. Testing with DNA from different pathogens, including bacteria and other protozoa, confirmed the high specificity of the assay. We propose the use of this PCR assay as an accurate, rapid, and effective diagnostic method for the detection and discrimination of these three morphologically indistinguishable Entamoeba species in both routine diagnosis of amoebiasis and epidemiological surveys.Infections with Entamoeba spp. can result in either a harmless colonization of the intestine or an invasion of the colon wall and damage of other host tissues such as the liver, lung, and brain (amoebiasis). In most cases, a clinical diagnosis of amoebiasis can be confirmed and usually depends on the visualization of parasites by light microscopy of a wet smear or stained specimens. This procedure is inexpensive and simple, but it has several limitations, such as being incapable of distinguishing between the cysts and trophozoites of the diseasecausing species Entamoeba histolytica, the nonpathogenic species Entamoeba dispar, and the amphizoic amoeba Entamoeba moshkovskii, which occasionally infects humans. Multiple samples often have to be requested and examined, and the presence of cysts of different species of Entamoeba, Iodamoeba, or Endolimax can make the diagnosis even more difficult (4, 5). Furthermore, with the reports of sporadic cases of human infection with E. moshkovskii (3, 6) and the recent finding of a high prevalence and association of E. moshkovskii with E. histolytica and E. dispar in young children in Bangladesh (2), the differentiation of the three species in clinical samples by other means becomes of great importance both for diagnosis and for epidemiological studies.

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Section: Discussionmentioning

confidence: 99%

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Differential Detection of Entamoeba histolytica , Entamoeba dispar , and Entamoeba moshkovskii by a Single-Round PCR Assay

Hamzah¹,

et al. 2006

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“…QIAamp tissue kit spin columns (QIAGEN, Hilden, Germany) have been used for the purification of DNA from microscopy-positive samples stored at Ϫ20°C in phosphate-buffered saline (196,199) and for DNA isolation using other modifications (such as treatment with 2% polyvinylpolypyrrolidone [Sigma]) which improve the sensitivity of the PCR (197). The use of the QIAamp stool kit for the extraction of DNA from fecal samples was a major advance, and this has proven to be the most widely accepted method for DNA ex- (50,51,54,65,82,129,196,199) and are now used widely in clinical research laboratories in developed nations, as they minimize the extraction time and the DNA can be extracted directly from the feces without the need to culture the parasites.…”

Section: Methods Of Dna Extractionmentioning

confidence: 99%

Laboratory Diagnostic Techniques for Entamoeba Species

et al. 2007

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SUMMARY The genus Entamoeba contains many species, six of which (Entamoeba histolytica, Entamoeba dispar, Entamoeba moshkovskii, Entamoeba polecki, Entamoeba coli, and Entamoeba hartmanni) reside in the human intestinal lumen. Entamoeba histolytica is the causative agent of amebiasis and is considered a leading parasitic cause of death worldwide in humans. Although recent studies highlight the recovery of E. dispar and E. moshkovskii from patients with gastrointestinal symptoms, there is still no convincing evidence of a causal link between the presence of these two species and the symptoms of the host. New approaches to the identification of E. histolytica are based on detection of E. histolytica-specific antigen and DNA in stool and other clinical samples. Several molecular diagnostic tests, including conventional and real-time PCR, have been developed for the detection and differentiation of E. histolytica, E. dispar, and E. moshkovskii in clinical samples. The purpose of this review is to discuss different methods that exist for the identification of E. histolytica, E. dispar, and E. moshkovskii which are available to the clinical diagnostic laboratory. To address the need for a specific diagnostic test for amebiasis, a substantial amount of work has been carried out over the last decade in different parts of the world. The molecular diagnostic tests are increasingly being used for both clinical and research purposes. In order to minimize undue treatment of individuals infected with other species of Entamoeba such as E. dispar and E. moshkovskii, efforts have been made for specific diagnosis of E. histolytica infection and not to treat based simply on the microscopic examination of Entamoeba species in the stool. The incorporation of many new technologies into the diagnostic laboratory will lead to a better understanding of the public health problem and measures to control the disease.

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“…The DNA of the above mentioned species you can vary the amoebas by using molecular biology methods such as: nested PCR, real-time PCR, LC-PCR, PCR-SHELA, Reverse Line Hybridization Assay [8][9][10]29,38,39,41,[46][47][48][49]. These methods according to Tanyuksel et al, are characterized by high sensitivity (> 70%) and a very high specificity (> 90%) in the diagnosis of intestinal amoebiasis, parenteral amoebiasis and amoebic liver abscess [6].…”

Section: Cysts and Trophozoites Of Entamoeba Histolytica Microscopicamentioning

confidence: 99%

Entamoeba histolytica - Pathogenic Protozoan of the Large Intestine in Humans

Nowak¹

2015

J Clin Microbiol Biochem Technol

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Entamoeba histolytica is a cosmopolitan, parasitic protozoan of human large intestine, which is a causative agent of amoebiasis. Amoebiasis manifests with persistent diarrhea containing mucus or blood, accompanied by abdominal pain, flatulence, nausea and fever. In some cases amoebas may travel through the bloodstream from the intestine to the liver or to other organs, causing multiple abscesses. Amoebiasis is a dangerous, parasitic disease and after malaria the second cause of deaths related to parasitic infections worldwide. The highest rate of infections is observed among people living in or traveling through the tropics. Laboratory diagnosis of amoebiasis is quite difficult, comprising of microscopy and methods of molecular biology. Pathogenic species Entamoeba histolytica has to be differentiated from other nonpathogenic amoebas of the intestine, so called commensals, that very often live in the human large intestine and remain harmless. Other intestinal commensals are Entamoeba dispar and Entamoeba moshkovskii, morphologically the same as pathogenic species Entamoeba histolytica sensu stricto. The differential diagnosis of these three amoebas is possible with detection of their DNA.get sick, and 100 thousand people die because of amoebosis-induced infection dysentery [2].E. histolytica belongs to the cosmopolitan parasites, occurring throughout the globe. Particularly exposed to this parasite are people living and traveling to the tropical and subtropical zones (Asia, Africa, India, Indonesia, Mexico, South America, South Africa). These zones climatic conditions are optimal for the Protozoan cysts, which described the external environment can survive for many days. This fact contributes to the increase in the number of infections slider Entamoeba histolytica amongst people in tropical zones. According to Weinke to high-risk groups who are particularly vulnerable to being infected by E. histolytica include men with a homosexual orientation [3]. Prevalence, it means the incidence of infection is low in Poland and is about 1%, while in tropical countries can claim up to 50%. Most of the amoebiasis cases reported in Poland applies to people returning from a different climate zone and foreigners.The digestive tract may be inhabited by several species of nonpathogenic amoebas in the family Entamoebidae, which include Entamoeba coli, Entamoeba hartmanni, Entamoeba polecki, Endolimax nana,. The aforementioned amoebas are so called commensals of colon, which may settle the intestinal mucosa physiologically not attracting damages. There are only isolated reports in the literature that some commensals may lead to pathogenic actions in the human large intestine. There have been a couple of cases of diarrhea problems caused by infections of Endolimax nana, Entamoeba polecki and I. bűtschlii in immunocompromised patients [6].In 1925, Brumpt suggested that within the family Entamoebidae there are species Entamoeba dispar, which is morphologically Abbreviations E.

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Detection and Identification of Entamoeba Species in Stool Samples by a Reverse Line Hybridization Assay

Cited by 48 publications

References 12 publications

Differential Detection of Entamoeba histolytica , Entamoeba dispar , and Entamoeba moshkovskii by a Single-Round PCR Assay

Differential Detection of Entamoeba histolytica , Entamoeba dispar , and Entamoeba moshkovskii by a Single-Round PCR Assay

Laboratory Diagnostic Techniques for Entamoeba Species

Entamoeba histolytica - Pathogenic Protozoan of the Large Intestine in Humans

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