A critical assessment of two real-time PCR assays targeting the (SSU) rRNA and gdh genes for the molecular identification of<i>Giardia intestinalis</i>in a clinical laboratory

Boadi, Samuel; Polley, Spencer D.; Kilburn, Sally; Mills, Graham A.; Chiodini, Peter L.

doi:10.1136/jclinpath-2014-202224

Cited by 19 publications

(17 citation statements)

References 23 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…This makes PCR a more convenient method, as a single sample suffices and thus there is no longer a need for collecting stool samples on three consecutive days in order to reach a diagnostic sensitivity similar to microscopy. Also, because PCR detection does not rely on the detection of intact cysts or motile trophozoites, sensitive detection can still be achieved without the need of preservatives and/or different staining methods …”

Section: New Detection Methodsmentioning

confidence: 99%

“…Therefore, microscopy requires the availability of highly trained personnel and is very labor intensive . Failures to find and differentiate protozoa by microscope lead to decreased sensitivity that may result in an inadequate diagnosis of intestinal protozoa using microscopy . Even between European reference laboratories, substantial differences were found in detection rates and thus diagnosis of intestinal protozoa using microscopy .…”

Section: Traditional Detectionmentioning

confidence: 99%

“…When sensitivity of PCR was compared with microscopy, it was seen that parasitic DNA could still be amplified and detected in cases where a sample was negative for protozoa using microscopy . Possibly, DNA can still be found in stool samples when cysts concentration is too low for microscopic detection or when cysts are not apparent in stool samples.…”

Section: Clinical Issues When Requesting Polymerase Chain Reactionmentioning

confidence: 99%

“…Because sensitivity of different primers and probes varies substantially, so do detection rates between individual laboratories. Due to the heterogeneous implementation of different PCR assays, it is uncertain if sufficient detection rates are achieved in individual laboratories …”

Section: Clinical Issues When Requesting Polymerase Chain Reactionmentioning

confidence: 99%

See 3 more Smart Citations

Clinical consequences of polymerase chain reaction‐based diagnosis of intestinal parasitic infections

Rijsman

Monkelbaan

Kusters

2016

J of Gastro and Hepatol

View full text Add to dashboard Cite

The implementation of polymerase chain reaction (PCR)-based diagnostics of intestinal protozoa has led to higher sensitivity and (subtype) specificity, more convenient sampling, and the possibility for high-throughput screening. PCR for routine detection of human intestinal protozoa in fecal samples is used by an increasing number of clinical laboratories. This paper discusses the recent developments in the diagnosis of intestinal protozoa, with an emphasis on PCR-based diagnostics. Although many reviews have described the technical aspects of PCR-based diagnostics, this review focuses on the clinical consequences that result from the shift from microscopic toward PCR-based diagnostics. Despite its undisputed superiority, the use of PCR comes with challenges that clinicians should be aware of.

show abstract

Section: New Detection Methodsmentioning

confidence: 99%

Section: Traditional Detectionmentioning

confidence: 99%

Section: Clinical Issues When Requesting Polymerase Chain Reactionmentioning

confidence: 99%

Section: Clinical Issues When Requesting Polymerase Chain Reactionmentioning

confidence: 99%

See 2 more Smart Citations

Clinical consequences of polymerase chain reaction‐based diagnosis of intestinal parasitic infections

Rijsman

Monkelbaan

Kusters

2016

J of Gastro and Hepatol

View full text Add to dashboard Cite

show abstract

“…Regarding the real-time PCR on G. intestinalis, Bertrand et al showed that for an initial concentration of 180 cysts per 200 μl of preparation, only 33% of positive responses were observed [38,39]. Indeed, a relatively low sensitivity (93.4%) was observed by targeting sequences of the small subunit gene of rRNA [40] as in our study.…”

Section: Discussionmentioning

confidence: 41%

Evaluation of a Real-Time Polymerase Chain Reaction for the Laboratory Diagnosis of Giardia intestinalis in Stool Samples from Schoolchildren from the Centre-Ouest and Plateau Central Regions of Burkina Faso

Diagbouga¹,

Kientega²,

Erismann³

et al. 2017

Appli Micro Open Access

View full text Add to dashboard Cite

Objective: Giardiasis, a zoonotic, diarrhoeal disease with worldwide occurrence, is routinely diagnosed by microscopic examination of stool samples. However, implementation of this method relies on skilled personal, it is time consuming and relatively low in sensitivity. A superior diagnostic approach to detect the causative agent Giardia intestinalis would, hence, be highly desirable. The current study aimed to assess real-time polymerase chain reaction (PCR) for the detection of G. intestinalis as an alternative to microscopy. Material and methods:Stool samples from healthy schoolchildren, aged 8-14 years, from eight schools of the Centre-Ouest and Plateau Central regions in Burkina Faso were collected within a cross-sectional study in February 2015. Microscopic examination was performed on two faecal samples collected over two consecutive days from 441 schoolchildren. Each faecal specimen was examined using Kato-Katz and formol-ether methods of concentration in addition to direct examination. Real-time PCR was used to detect G. intestinalis in all microscopy-positive and a random sample of microscopy-negative samples.Results: Microscopic examination revealed 94 microscopy-positive samples, and an overall G. intestinalis prevalence of 27.2%. Using the microscopic examination as the 'gold' standard, the overall sensitivity of real-time PCR was demonstrated to be 76.6% ranging from 58.3% to 94.1% and the specificity was 96.2% ranging from 96.2% to 100% across schools. Conclusion:Real-time PCR appears to be a solid detection method for G. intestinalis in the current setting. However, it needs to be further optimized to become a more sensitive tool for G. intestinalis diagnosis in low-income settings.

show abstract

Bowel Histology of CVID Patients Reveals Distinct Patterns of Mucosal Inflammation

et al. 2021

View full text Add to dashboard Cite

Diarrhea is the commonest gastrointestinal symptom in patients with common variable immunodeficiency (CVID). Different pathologies in patients’ bowel biopsies have been described and links with infections have been demonstrated. The aim of this study was to analyze the bowel histology of CVID patients in the Royal-Free-Hospital (RFH) London CVID cohort. Ninety-five bowel histology samples from 44 adult CVID patients were reviewed and grouped by histological patterns. Reasons for endoscopy and possible causative infections were recorded. Lymphocyte phenotyping results were compared between patients with different histological features. There was no distinctive feature that occurred in most diarrhea patients. Out of 44 patients (95 biopsies), 38 lacked plasma cells. In 14 of 21 patients with nodular lymphoid hyperplasia (NLH), this was the only visible pathology. In two patients, an infection with Giardia lamblia was associated with NLH. An IBD-like picture was seen in two patients. A coeliac-like picture was found in six patients, four of these had norovirus. NLH as well as inflammation often occurred as single features. There was no difference in blood lymphocyte phenotyping results comparing groups of histological features. We suggest that bowel histology in CVID patients with abdominal symptoms falls into three major histological patterns: (i) a coeliac-like histology, (ii) IBD-like changes, and (iii) NLH. Most patients, but remarkably not all, lacked plasma cells. CVID patients with diarrhea may have an altered bowel histology due to poorly understood and likely diverse immune-mediated mechanisms, occasionally driven by infections.

show abstract

A critical assessment of two real-time PCR assays targeting the (SSU) rRNA and gdh genes for the molecular identification ofGiardia intestinalisin a clinical laboratory

Abstract: The Verweij real-time PCR was robust and the most sensitive assay suited for use as a first-line diagnostic test for giardiasis.

Cited by 19 publications

References 23 publications

Clinical consequences of polymerase chain reaction‐based diagnosis of intestinal parasitic infections

Clinical consequences of polymerase chain reaction‐based diagnosis of intestinal parasitic infections

Evaluation of a Real-Time Polymerase Chain Reaction for the Laboratory Diagnosis of Giardia intestinalis in Stool Samples from Schoolchildren from the Centre-Ouest and Plateau Central Regions of Burkina Faso

Bowel Histology of CVID Patients Reveals Distinct Patterns of Mucosal Inflammation

Contact Info

Product

Resources

About