Real-time PCR for the detection of Dientamoeba fragilis in fecal samples

Verweij, Jaco J.; Mulder, Bert; Poell, Bregje; Middelkoop, Dorien van; Brienen, Eric A. T.; Lieshout, Lisette van

doi:10.1016/j.mcp.2007.05.006

Cited by 99 publications

(83 citation statements)

References 21 publications

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“…A selection bias on the patient population cannot be ruled out, because we have no log from primary care showing to which extent all eligible patients were referred for inclusion. We used PCR for the detection of D. fragilis, which is known to be a sensitive method for detecting the parasite, 42 and we analyzed three consecutive samples thereby overcoming the problem with intermittent shedding of the parasite in feces. 43 Our eradication rate of 60% of MZ on D. fragilis is consistent with the literature 24,[43][44][45][46][47] supporting the validity of our estimate of microbiological response.…”

Section: Discussionmentioning

confidence: 99%

Treatment of Dientamoeba fragilis in Patients with Irritable Bowel Syndrome

Engsbro

Stensvold²,

Nielsen³

et al. 2012

The American Journal of Tropical Medicine and Hygiene

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Abstract. The role of Dientamoeba fragilis in irritable bowel syndrome (IBS) is incompletely known. We aimed to investigate whether eradication of D. fragilis alleviates symptoms in IBS. Twenty-five D. fragilis-positive IBS patients were treated with Metronidazole (MZ) or Tetracycline. The patients were mostly female (89%), and mean age (SD) was 35.1 (8.2) years. Microbiological response, evaluated 2 weeks post-treatment, was observed in 15 of 25 patients (60%), all by MZ. Clinical response, defined as adequate relief of symptoms, was observed in 7 of 22 patients (32%), all by MZ. In a logistic regression analysis, we found no significant association between clinical and microbiological response. This case study did not support our hypothesis of a simple association between D. fragilis and IBS. Some D. fragilis-infections were insufficiently treated by MZ. Further studies into the prevalence and effect of eradication of D. fragilis in IBS and into efficient treatments of D. fragilis are warranted.

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Section: Discussionmentioning

confidence: 99%

Treatment of Dientamoeba fragilis in Patients with Irritable Bowel Syndrome

Engsbro

Stensvold²,

Nielsen³

et al. 2012

The American Journal of Tropical Medicine and Hygiene

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“…After homogenization in stool lysis buffer, 200 mg sterile zirconia/ silica beads (diameter 0.01 mm; BioSpec Products, Roth, Karlsruhe, Germany) were added for a mechanical disruption procedure of 6 min and 30 Hz in a TissueLyzer (Qiagen Retsch GmbH, Hannover, Germany), followed by lysis for 5 min at 95°C. Real-time PCR was used to detect D. fragilis (14), and the results are presented as positive or negative. For positive samples, C T values were recorded.…”

Section: Methodsmentioning

confidence: 99%

“…Using real-time PCR (14), DNA of D. fragilis was detected in the majority (516/688 [75.0%]) of the stool Age was a significant factor and acted as a confounder in both final multivariable models. The final multivariable logistic regression model for testing positive for D. fragilis had two risk factors: being Ͼ3 years old and having a history of traveling abroad during the previous 2 months, with odds ratios of 3.49 (95% CI, 1.59 to 7.70) and 3.15 (95% CI, 1.06 to 9.36), respectively.…”

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confidence: 99%

Dientamoeba fragilis, a Commensal in Children in Danish Day Care Centers

et al. 2017

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Dientamoeba fragilis is an intestinal protozoan of debated clinical significance. Here, we present cross-sectional and longitudinal observations on D. fragilis in children aged 0 to 6 years from a 1-year multi-day-care-center cohort study set in Copenhagen, Denmark. The inclusion period for the cohort was 2009 through 2012. Stool samples collected from the children were accompanied by questionnaires completed by the parents or guardians of the children. Using real-time PCR, D. fragilis was detected in the first stool sample from 97 of 142 (68.3%) children. We evaluated the associations between seven plausible risk factors (age, sex, having siblings, having domestic animals at home, having had infant colic, recent history of intake of antibiotics, and recent history of travel abroad) as well as six reported symptoms (lack of appetite, nausea, vomiting, abdominal pain, weight loss, and diarrhea) and testing positive for D. fragilis. The final multivariable model identified being Ͼ3 years old and having a history of recent travel abroad as risk factors for testing positive for D. fragilis. Moreover, univariable analyses indicated that having siblings was a risk factor. There was no statistical association between a recent history of gastrointestinal symptoms and testing positive for D. fragilis. Among the 108 children who were represented by Ն2 samples and thus included in the longitudinal analysis, 32 tested negative on the first sample and positive later, and the last sample from each of the 108 children was positive. The results are in support of D. fragilis being a common enteric commensal in this population.KEYWORDS Denmark, dientamoebiasis, infants, preschoolers, risk factors, symptoms, toddlers D ientamoeba fragilis is an intestinal protozoan of debated clinical significance (1-7). In Denmark, D. fragilis appears to be common in the apparently healthy adult population (8); meanwhile, baseline data on the prevalence in apparently healthy children are lacking. Dientamoeba fragilis is commonly detected in stools from children tested for gastrointestinal pathogens, with a peak (ca. 70% of samples positive) in 7-year-olds (9). A randomized placebo-controlled clinical trial failed to show improved clinical outcome in D. fragilis-positive children treated with metronidazole compared with those treated with a placebo, and the observed eradicative effect of metronidazole was moderate and transient, and a spontaneous decrease in the proportion of children testing positive was observed in the placebo group (10).We recently established an open cohort of children aged 0 to 6 years attending municipal day care centers in Copenhagen, Denmark, with a primary research focus on investigating enteroaggregative Escherichia coli over time (11-13). Using real-time PCR (14), DNA of D. fragilis was detected in the majority (516/688 [75.0%]) of the stool

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“…A real-time PCR targeting 77 bp of the SSU rRNA gene was the first molecular assay developed as a screening tool, potentially replacing expensive and time-consuming parasitological diagnosis (127). Interestingly, with a real-time PCR targeting the 5.8S ribosomal gene, which also included an internal process control, it was shown that no reduction in amplification efficiency could be detected when comparing fresh material with material that had been stored unpreserved at 4°C for 8 weeks (128).…”

Section: Dientamoeba Fragilismentioning

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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Real-time PCR for the detection of Dientamoeba fragilis in fecal samples

Cited by 99 publications

References 21 publications

Treatment of Dientamoeba fragilis in Patients with Irritable Bowel Syndrome

Treatment of Dientamoeba fragilis in Patients with Irritable Bowel Syndrome

Dientamoeba fragilis, a Commensal in Children in Danish Day Care Centers

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

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