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Pathogens (Basel, Switzerland) Jun 2023We aimed to assess the performance of the Novodiag Stool Parasites (NSP) assay in the diagnosis of the most common intestinal protozoan and microsporidia infections.
OBJECTIVES
We aimed to assess the performance of the Novodiag Stool Parasites (NSP) assay in the diagnosis of the most common intestinal protozoan and microsporidia infections.
METHODS
A panel of 167 selected stool samples was retrospectively analysed with the NSP assay and compared to routine microscopy and qPCR methods for the detection of pathogenic protozoa and microsporidia.
RESULTS
Whereas specificity was high for all protozoa and microsporidia, NSP sensitivity was strongly dependent on the comparative method used as reference. When compared to microscopic methods, NSP sensitivity was high (96.7 to 100%) for , and but was lower for (85.2%) and ≤50% for and . In comparison to conventional qPCR, the NSP assay demonstrated lower sensitivity characteristics dependent on parasite loads, reaching 60 to 70% for , , spp. and Sensitivity was 100% for , but none of the five samples containing spp. were detected.
CONCLUSIONS
The overall performance of the NSP assay in the diagnosis of gastrointestinal protozoa and microsporidia seems to be better than or equivalent to that observed with microscopic methods but inferior to that obtainable with classical targeted qPCR.
PubMed: 37513736
DOI: 10.3390/pathogens12070889 -
Pathogens (Basel, Switzerland) Oct 2022The enteric protozoan parasites spp., and are-to various extents-contributors to the burden of gastrointestinal illness in high-income countries. Detection of these...
The enteric protozoan parasites spp., and are-to various extents-contributors to the burden of gastrointestinal illness in high-income countries. Detection of these pathogens by microscopy examination is challenging because of the limited sensitivity and need for specific staining procedures. We developed and optimised a new multiplex real-time PCR assay for the simultaneous detection of spp., and in clinical (stool) samples. The diagnostic performance of the assay was evaluated against a large panel of well-characterised DNA samples positive for spp. ( = 126), ( = 132) and ( = 49). The specificity of the test was assessed against a DNA panel from other intestinal or phylogenetically related parasites ( = 105) and faecal DNA from individuals without clinical manifestations ( = 12). The assay exhibited a diagnostic sensitivity of 0.90-0.97 and a diagnostic specificity of 1. The limit of detection was estimated for (1 oocyst) and (5 × 10 cysts). The method allowed the detection of four species (, , and ) and five assemblages (A-E) without cross-reacting with other parasites belonging to the phyla Amoebozoa, Apicomplexa, Euglenozoa, Microsporidia, Nematoda and Platyhelminthes. This newly developed multiplex real-time PCR assay represents a novel alternative for the rapid and accurate detection of , and in clinical settings.
PubMed: 36365028
DOI: 10.3390/pathogens11111277 -
Parasitology International Feb 2024Dientamoeba fragilis (D. fragilis) represents a common protozoan in both high and low income countries. Despite this, epidemiological data on dientamoebiasis are still...
Dientamoeba fragilis (D. fragilis) represents a common protozoan in both high and low income countries. Despite this, epidemiological data on dientamoebiasis are still limited, and it is possible that the actual prevalence rates of D. fragilis have been underestimated due to the challenges in its detection and identification. In the present study, symptomatic patients from Rome (Central Italy) were surveyed for two years to determine D. fragilis percentage of infection and genotypes. Stool samples collection was performed over 864 patients, DNA extracted, and RT-PCR performed by the SeeGene Allplex™ Gastrointestinal Parasite Panel Assays. Seventy-nine resulted positive for D. fragilis (9.1%). Co-infections were detected in 22 isolates: 21 displayed Blastocystis sp. + D. fragilis (27.8%). Based on the sequence of a central fragment of the SSU rRNA gene, only genotype 1 was identified. These findings are among the few available data regarding genetic diversity of D. fragilis in Italy. Large-scale human and animal research are required to enhance our knowledge of prevalence, host range, genetic variability and zoonotic transmission of this little-known intestinal protozoan.
Topics: Animals; Humans; Dientamoeba; Genotype; Intestinal Diseases, Parasitic; Dientamoebiasis; Feces; Italy
PubMed: 37838287
DOI: 10.1016/j.parint.2023.102816 -
International Maritime Health 2023Poland has experienced increased economic migration since 2021. Large waves of migrants, mostly from Asian and African countries, are trying to get into the European...
BACKGROUND
Poland has experienced increased economic migration since 2021. Large waves of migrants, mostly from Asian and African countries, are trying to get into the European Union by crossing Poland's eastern border illegally. The influx of illegal migrants into Poland is the result of a policy adopted by the Belarusian and Russian regimes that are trying to provoke another migrant crisis in Europe. In the opinion of some Polish politicians illegal migration contributes to the spread of parasitic diseases in our country as many migrants arriving into Poland carry intestinal parasites. The aim of this study was to assess the prevalence of infections with intestinal parasites in the Polish Border Guard officers safeguarding Poland's eastern borders.
MATERIALS AND METHODS
Parasitological diagnostics was carried out between April and May 2023. The study involved 218 Polish Border Guard officers from the Podlaski Border Guard Unit (PBGU) and 209 officers from the Bug Border Guard Unit (BBGU), whose task is to patrol and safeguard Poland's border with Ukraine and Belarus. Faecal examinations were performed using three different light microscopy testing methods (direct smear, decantation, flotation) at the Department of Epidemiology and Tropical Medicine at the Military Institute of Medicine - National Research Institute, Warsaw, Poland.
RESULTS
Considered to be potentially pathogenic intestinal parasites were diagnosed in 20 out of 218 officers serving in the PBGU (8.7% infected with Blastocystis spp., 0.5% with Dientamoeba fragilis) and in 9 out of 209 officers serving in the BBGU (3.8% infected with Blastocystis spp., 0.5% with Dientamoeba fragilis). There were no infections with nematodes, cestodes or trematodes in the study participants. No correlation was found between a parasitic infection and the presence of diarrhoea or other gastrointestinal symptoms within 6 months prior to the study in both groups.
CONCLUSIONS
Although Polish Border Guard officers deployed to the eastern border are exposed to difficult environmental conditions and have frequent contacts (either directly or indirectly) with migrants arriving from countries which report high incidence of parasitic infections, the rates of infections with potentially pathogenic protozoa in officers from the PBGU and BBGU are low and mainly attributable to pathogens which are widespread in the general Polish population. Low rates of parasitic infections in officers serving in the border zone suggest that the epidemiological situation of parasitic diseases in East Poland is satisfactory and that the disease prevention strategies (including the use of personal protection gear) implemented by the Polish medical services are effective.
Topics: Humans; Poland; Intestinal Diseases, Parasitic; European Union; Military Personnel; Europe
PubMed: 37781943
DOI: 10.5603/imh.97185 -
Microorganisms Oct 2023Indigenous people live in remote areas of Colombia. Multiple infections with bacteria, protozoa and/or helminths are common, as well as colonization in various forms....
Indigenous people live in remote areas of Colombia. Multiple infections with bacteria, protozoa and/or helminths are common, as well as colonization in various forms. This study focused on the question of whether and to what extent various pathogens interact with each other. Therefore, a mathematical approach was retrospectively applied to PCR-based data of 244 stool samples, collected in two datasets. A stable cluster solution of the pathogens assessed was determined, and a unique configuration between / spp./ forming cluster 1 and was verified. A pathogen density-dependent interplay appeared between the / spp./ cluster, and . The applied mathematical approach demonstrated that co-infections with parasites of questionable pathological relevance such as and can be of diagnostic relevance due to their ability to promote or repress other pathogens. With the increasing availability of highly sensitive multiplexed molecular diagnostic approaches even in resource-limited settings, where multiple colonization of infection events with enteric pathogens in parallel are common, the importance of interpreting whole pathogen patterns rather than just individual pathogen detection may become more and more relevant.
PubMed: 38004654
DOI: 10.3390/microorganisms11112642 -
International Journal of Molecular... Aug 2023Globally, over 3.5 billion people are infected with intestinal parasites each year, resulting in over 200,000 deaths. Three of the most common protozoan pathogens that...
Globally, over 3.5 billion people are infected with intestinal parasites each year, resulting in over 200,000 deaths. Three of the most common protozoan pathogens that affect the gastrointestinal tract of humans are spp., , and . Other protozoan agents that have been implicated in gastroenteritis in humans include , , , and the microsporidia and . Genetic Signatures previously developed a 3base™ multiplexed Real-Time PCR (mRT-PCR) enteric protozoan kit (EP001) for the detection of //, spp., , . , and . We now describe improvements to this kit to produce a more comprehensive assay, including , , and , termed EP005. The clinical performance of EP005 was assessed using a set of 380 clinical samples against a commercially available PCR test and other in-house nucleic acid amplification tests where commercial tests were not available. All methods provided at least 90% agreement. EP005 had no cross-reactivity against 82 organisms commonly found in the gut. The EP005 method streamlines the detection of gastrointestinal parasites and addresses the many challenges of traditional microscopic detection, resulting in cost savings and significant improvements in patient care.
Topics: Humans; Cryptosporidiosis; Cryptosporidium; Gastrointestinal Diseases; Communicable Diseases; Protozoan Infections; Giardia lamblia
PubMed: 37686192
DOI: 10.3390/ijms241713387 -
Nutrients Aug 2022Obesity is an epidemic causing a metabolic health crisis. Herein, the interactions between the gut prokaryotic and eukaryotic communities, metabolic comorbidities and...
Obesity is an epidemic causing a metabolic health crisis. Herein, the interactions between the gut prokaryotic and eukaryotic communities, metabolic comorbidities and diet were studied. Stool samples from 56 subjects, 47 with type III obesity and 9 with type II obesity and cardiovascular risk or metabolic disease, were assessed for the richness, diversity and ecology of the bacterial gut community through metagenomics, together with the study of the presence of common unicellular eukaryote parasites ( sp., and ) by qPCR. Clinical information regarding metabolic comorbidities and non-alcoholic hepatic fatty liver disease was gathered. To assess the quality of the patients' diet, each participant filled in three dietary questionnaires. The most prevalent parasite sp. (46.4%), together with (8.9%), was found to be associated with higher mean diversity indexes regarding non-colonized subjects; the opposite of that which was observed in those with (16.1%). In terms of phyla relative abundance, with sp. and , very slight differences were observed; on the contrary, was related to an increase in Bacteroidetes and Proteobacteria, and a decrease in Firmicutes and Actinobacteria, presenting the lowest Firmicutes/Bacteroidetes ratio. At genus level, sp. and/or was accompanied with an increase in spp., and a decrease in spp., spp. and spp., while was associated with an increase in spp., and a decrease in spp., spp. and spp., and the highest spp./ spp. ratio. Participants with non-alcoholic hepatic fatty liver presented a higher Firmicutes/Bacteroidetes ratio, and those with type 2 diabetes displayed a significantly lower spp./ spp. ratio, due to an overrepresentation of the genus spp. The presence of parasites was associated with variations in the richness, diversity and distribution of taxa in bacterial communities, confirming a gain in diversity associated with sp. and providing different functioning of the microbiota with a potential positive effect on comorbidities such as type 2 diabetes, insulin resistance and metabolic syndrome. Future basic and clinical studies should assess the beneficial or pathogenic effect of these eukaryotes on obese subjects and focus on deciphering whether they may imply a healthier metabolic profile.
Topics: Animals; Bacteria; Blastocystis; Diabetes Mellitus, Type 2; Feces; Humans; Obesity, Morbid; Parasites
PubMed: 35956387
DOI: 10.3390/nu14153211 -
Journal of Pediatric Gastroenterology... Dec 2021This survey was undertaken to obtain insight in the attitude of Dutch physicians towards pathogenicity, diagnostic- and therapeutic approach towards Dientamoeba fragilis...
This survey was undertaken to obtain insight in the attitude of Dutch physicians towards pathogenicity, diagnostic- and therapeutic approach towards Dientamoeba fragilis in children. Physicians were invited by e-mail for a questionnaire. A total of 211 of 450 physicians (46.9%) completed the questionnaire, including 67 general practitioners (GPs) and 144 pediatricians. Of all respondents, 175 of 211 (82.9%) considered D fragilis a "potential pathogen", when other causes of gastro-intestinal complaints are ruled out. Only 16 of 211 (7.6%) performed diagnostic tests regularly. Diagnostic tests were performed by 162 of 211 (77%) of respondents in children with diarrhea and abdominal pain in consideration of duration of symptoms. Fecal polymerase chain reaction (PCR) was diagnostic modality of preference. Eighty-nine of 142 (62.7%) prescribed metronidazole as antibiotic of first choice. This study shows heterogeneity in clinical practice amongst Dutch physicians regarding diagnostic- and therapeutic approach of D fragilis in children. Different attitude towards pathogenicity and inconsistent guidelines could be causative factors.
Topics: Child; Dientamoeba; Dientamoebiasis; Feces; General Practitioners; Humans; Netherlands; Pediatricians; Surveys and Questionnaires
PubMed: 34520404
DOI: 10.1097/MPG.0000000000003297 -
Euro Surveillance : Bulletin Europeen... Jul 2019BackgroundDespite the global distribution of the intestinal protozoan its clinical picture remains unclear. This results from underdiagnosis: microscopic screening...
BackgroundDespite the global distribution of the intestinal protozoan its clinical picture remains unclear. This results from underdiagnosis: microscopic screening methods either lack sensitivity (wet preparation) or fail to reveal (formalin-fixed sample).AimIn a retrospective study setting, we characterised the clinical picture of dientamoebiasis and compared it with giardiasis. In addition, we evaluated an improved approach to formalin-fixed samples for suitability in diagnostics.MethodsThis study comprised four parts: (i) a descriptive part scrutinising rates of findings; (ii) a methodological part analysing an approach to detect -like structures in formalin samples; (iii) a clinical part comparing demographics and symptoms between patients with dientamoebiasis (n = 352) and giardiasis (n = 272), and (iv) a therapeutic part (n = 89 patients) investigating correlation between faecal eradication and clinical improvement.ResultsThe rate of findings increased 20-fold after introducing criteria for -like structures in formalin-fixed samples (88.9% sensitivity and 83.3% specificity). A further increase was seen after implementing faecal PCR. Compared with patients with giardiasis, the symptoms in the group lasted longer and more often included abdominal pain, cramping, faecal urgency and loose rather than watery stools. Resolved symptoms correlated with successful faecal eradication (p < 0.001).ConclusionsPreviously underdiagnosed, has become the most frequently recorded pathogenic enteroparasite in Finland. This presumably results from improved diagnostics with either PCR or detection of -like structures in formalin-fixed samples, an approach applicable also in resource-poor settings. Symptoms of dientamoebiasis differ slightly from those of giardiasis; patients with distressing symptoms require treatment.
Topics: Abdominal Pain; Adult; Animals; Diarrhea; Dientamoeba; Dientamoebiasis; Feces; Female; Finland; Giardiasis; Humans; Male; Middle Aged; Polymerase Chain Reaction; Retrospective Studies; Sex Distribution
PubMed: 31339096
DOI: 10.2807/1560-7917.ES.2019.24.29.1800546 -
Parasitology International Jun 2022Intestinal parasites are considered a growing public health problem, being protozoa the main cause of intestinal disease. The objective of our study is to compare the... (Observational Study)
Observational Study
INTRODUCTION
Intestinal parasites are considered a growing public health problem, being protozoa the main cause of intestinal disease. The objective of our study is to compare the detection of intestinal protozoa by microscopy versus real-time PCR, as well as to determine the most prevalent protozoa in our environment in the paediatric population.
METHOD
An observational longitudinal study was carried out, both by microscopy and real time-PCR in stool samples from children (0- 15 years) received from April 2019 to March 2021.Children were classified in two groups according if they had or not had clinical parasitosis. Microscopic examination was performed in all samples using the Ritchie concentration technique with the commercial Mini PARASEP system (Movaco-Grifols®). The presence of Cryptosporidium sp. was evaluated with the modified Ziehl-Neelsen acid-fast stain. The real-time PCR was performed to all samples using the Allplex ™ gastrointestinal parasite panel 4 (Seegene®).
RESULTS
During the study period, 500 samples were received, being positive 31 (6.2%) by microscopy and 256 (51.2 %) by PCR. By microscopy, Blastocystis hominis was the most frequently observed (4.8%), followed by Giardia lamblia (1.6%), Dientamoeba fragilis (0.2%) and Cryptosporidium species (0.2%). Regarding the identification by PCR, D. fragilis (35.2%) was mainly identified, followed by B. hominis (28.1%), G. lamblia (7%) and Cryptosporidium sp. (0.8%) without finding clear differences in aetiology according to age. In the case of B. hominis and D. fragilis, there were not differences in the detection of these protozoa between the control group and children with clinical parasitosis (p = 0.11).
CONCLUSIONS
Real-time PCR increases the detection of intestinal protozoa, being underdiagnosed by microscopy, especially D. fragilis, in which PCR is considered the most appropriate method for its detection.
Topics: Child; Cryptosporidiosis; Cryptosporidium; Entamoeba histolytica; Feces; Giardia lamblia; Humans; Intestinal Diseases, Parasitic; Longitudinal Studies; Real-Time Polymerase Chain Reaction
PubMed: 35149177
DOI: 10.1016/j.parint.2022.102558