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Clinical Microbiology and Infection :... Jan 2024The aim of this study was to assess the clinical significance of Dientamoeba fragilis (DF) and Blastocystis species (Bs) in human stool. (Observational Study)
Observational Study
OBJECTIVES
The aim of this study was to assess the clinical significance of Dientamoeba fragilis (DF) and Blastocystis species (Bs) in human stool.
METHODS
Observational study of patients ≥18 years, who were tested by stool multiplex PCR for bacteria and parasites between April 2019 and March 2022. Although DF and Bs are part of the PCR kit, these results are not routinely reported to the patient or the ordering physician. The main outcomes were the incidence of symptoms during 14 days before the referral to stool PCR test, and the incidence of several clinical outcomes during 60 days after the PCR test (symptoms, referrals to further evaluation, prescription of symptomatic, or antibiotic treatment).
RESULTS
A total of 27 918 patients were tested by stool PCR during the 3 study years. A total of 6215 (22.3%) and 5337 (19.2%) were positive for DF and Bs, respectively. The incidence of symptoms before the test was similar in those positive for Bs or DF and those with all-negative PCR (adjusted OR and 95% CI of 0.87 [0.80-0.95] and 0.82 [0.76-0.88] for Bs and DF, respectively), whereas significantly higher (2.47 [2.23-2.73]) in those positive for the other multiplex PCR assay components. During the 60 days after the test, the prevalence of any of the outcomes was similar in those positive for Bs or DF and those with negative PCR (adjusted OR and 95% CI of 0.92 [0.83-1.02] and 0.89 [0.81-0.97] for symptoms, 0.84 [0.75-0.94] and 0.93 [0.85-1.01] for referrals, 0.88 [0.75-1.03] and 0.82 [0.71-0.94] for symptomatic treatment, and 0.88 [0.75-1.02] and 0.86 [0.75-0.98] for antibiotic treatment in the Bs and DF positive individuals, respectively). The PCR cycle threshold was not associated with any of the outcomes.
DISCUSSION
Positive stool PCR for DF or Bs was not associated with any of the measured clinical outcomes.
Topics: Humans; Blastocystis; Dientamoeba; Clinical Relevance; Retrospective Studies; Multiplex Polymerase Chain Reaction; Feces; Anti-Bacterial Agents
PubMed: 37689266
DOI: 10.1016/j.cmi.2023.09.003 -
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 -
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 -
Dientamoeba fragilis associated with microbiome diversity changes in acute gastroenteritis patients.Parasitology International Dec 2023This study examined the correlation between intestinal protozoans and the bacterial microbiome in faecal samples collected from 463 patients in New Zealand who were...
This study examined the correlation between intestinal protozoans and the bacterial microbiome in faecal samples collected from 463 patients in New Zealand who were diagnosed with gastroenteritis. In comparison to traditional microscopic diagnosis methods, Multiplexed-tandem PCR proved to be more effective in detecting intestinal parasites. Among the identified protozoans, Blastocystis sp. and Dientamoeba fragilis were the most prevalent. Notably, D. fragilis was significantly associated with an increase in the alpha-diversity of host prokaryotic microbes. Although the exact role of Blastocystis sp. and D. fragilis as the primary cause of gastroenteritis remains debatable, our data indicates a substantial correlation between these protozoans and the prokaryote microbiome of their hosts, particularly when compared to other protists or patients with gastroenteritis but no detectable parasitic cause. These findings underscore the significance of comprehending the contributions of intestinal protozoans, specifically D. fragilis, to the development of gastroenteritis and their potential implications for disease management.
Topics: Animals; Humans; Dientamoeba; Intestinal Diseases, Parasitic; Blastocystis; Gastroenteritis; Parasites; Feces
PubMed: 37482266
DOI: 10.1016/j.parint.2023.102788 -
Zhongguo Xue Xi Chong Bing Fang Zhi Za... Jun 2023To investigate the prevalence of parasitic infections in human stool samples from a hospital in Chenzhou City, Hunan Province, so as to provide insights into the...
OBJECTIVE
To investigate the prevalence of parasitic infections in human stool samples from a hospital in Chenzhou City, Hunan Province, so as to provide insights into the management of intestinal parasitic diseases.
METHODS
Stool samples were collected from patients admitted to a hospital in Chenzhou City from September 2020 to March 2021, subjected to physiological saline smearing and microscopy for detection of intestinal parasites. The prevalence of parasitic infections and the species of parasites were descriptively analyzed.
RESULTS
The overall prevalence of intestinal parasitic infections was 1.61% in the 10 728 stool samples, and there were 3 samples with mixed infections of two parasite species. A total of seven parasite species were identified, including (162 cases, 1.55%), (5 cases, 0.05%), (5 cases, 0.05%), (one case, 0.01%), (one case, 0.01%), (one case, 0.01%) and (one case, 0.01%). The prevalence of intestinal parasitic infection was significantly higher among women than in men (2.14% vs. 1.25%; χ = 13.01, < 0.01), and a high prevalence rate was seen among patients at ages of 20 to 30 years (2.99%) and 80 years and older (2.86%); however, no age-specific prevalence of intestinal parasitic infection was detected (χ = 12.45, > 0.05).
CONCLUSIONS
The overall prevalence of intestinal parasitic infection was low among patients admitted to a hospital in Chenzhou City, and gender-specific prevalence was found. Food-borne and opportunistic parasites were predominant intestinal parasites, including , and .
Topics: Animals; Female; Humans; Male; Feces; Hospitals; Intestinal Diseases, Parasitic; Parasites; Prevalence; Young Adult; Adult; Aged, 80 and over
PubMed: 37455102
DOI: 10.16250/j.32.1374.2022211 -
Parasitology Research Jun 2023Intestinal parasitic infections are a global health problem that causes morbidity and mortality, especially in children living in rural areas. In this study, stool...
Intestinal parasitic infections are a global health problem that causes morbidity and mortality, especially in children living in rural areas. In this study, stool samples of pediatric patients with gastrointestinal complaints were examined by conventional and molecular methods to determine the prevalence of intestinal parasites. A total of 100 pediatric patients with gastrointestinal complaints and 50 healthy children were included in the study. Stool samples were collected from each child and examined by direct microscopic examination (native-Lugol method), formol-ethyl acetate concentration technique, Kinyoun's acid-fast staining, and Wheatley trichrome staining methods. Real-time PCR was used for the detection of Blastocystis spp. and D. fragilis in the stool samples. Sanger sequencing was used to identify Blastocystis spp. subtypes. One or more intestinal parasites were found in 12% (n = 100) of the patient group and 1% (n = 50) of the control group using conventional techniques. By using real-time PCR, Blastocystis spp. was discovered in 14% (14/100) of the patient group and 8% (4/50) of the control group. There was no significant difference in the frequency of Blastocystis spp. between the two groups. The most prevalent Blastocystis subtype was ST1 and the most frequent allele was a2 among the samples successfully amplified and sequenced. D. fragilis was detected in 17% (17/100) of the patient group and 8% (4/50) of the control group by real-time PCR. The prevalence of D. fragilis was not significantly different between the patient and control groups, as well. Blastocystis spp. and D. fragilis were found in high prevalence in pediatric patients with gastrointestinal complaints in this study. Although the role of these protists as a pathogen in humans is still controversial, it is supposed to the presence of the parasites are associated with gastrointestinal disorders such as diarrhea, abdominal pain, nausea, and vomiting. More case-control studies are needed to understand the pathogenic or commensal role of these parasites on the intestinal microbiota, especially in both patients with gastrointestinal disorders and healthy individuals.
Topics: Animals; Humans; Child; Parasites; Blastocystis Infections; Feces; Intestinal Diseases, Parasitic; Blastocystis; Gastrointestinal Diseases; Prevalence
PubMed: 37036521
DOI: 10.1007/s00436-023-07836-0 -
Tropical Parasitology 2022() diagnosis is an intestinal protozoan parasite globally found in rural and urban areas and is attracting a growing interest. Its prevalence in stool varies from 0.2%...
INTRODUCTION
() diagnosis is an intestinal protozoan parasite globally found in rural and urban areas and is attracting a growing interest. Its prevalence in stool varies from 0.2% to more than 19% depending upon the population studied.
MATERIALS AND METHODS
This study was based on the examination of 100 stool samples of randomly referred cases in a rural area in Motobus district, Kafr El-Sheikh governorate, Egypt. Our aim was to investigate the presence of in stool of the examined individuals using conventional polymerase chain reaction (PCR) compared to wet mount and trichrome stain with confirmation of infection by transmission electron microscopy.
RESULTS
was detected in 13/100 of the stool samples examined using wet mount smears, while trichrome stain detected 17/100. Conventional PCR diagnosed 41 cases of in the studied group. A very good agreement was found between wet mount and trichrome stain for diagnosing , while there was fair agreement between conventional PCR and both microscopy methods. Transmission electron microscope was performed on pooled positive samples that revealed the internal structures of trophozoite with its characteristic nucleus.
CONCLUSIONS
PCR technique was superior to microscopy for the detection of . Trichrome stain remains vital for microscopic diagnosis.
PubMed: 36643979
DOI: 10.4103/tp.tp_93_21 -
Biology Nov 2022The human gastrointestinal microbiota contains a diverse consortium of microbes, including bacteria, protozoa, viruses, and fungi. Through millennia of co-evolution, the... (Review)
Review
The human gastrointestinal microbiota contains a diverse consortium of microbes, including bacteria, protozoa, viruses, and fungi. Through millennia of co-evolution, the host-microbiota interactions have shaped the immune system to both tolerate and maintain the symbiotic relationship with commensal microbiota, while exerting protective responses against invading pathogens. Microbiome research is dominated by studies describing the impact of prokaryotic bacteria on gut immunity with a limited understanding of their relationship with other integral microbiota constituents. However, converging evidence shows that eukaryotic organisms, such as commensal protozoa, can play an important role in modulating intestinal immune responses as well as influencing the overall health of the host. The presence of several protozoa species has recently been shown to be a common occurrence in healthy populations worldwide, suggesting that many of these are commensals rather than invading pathogens. This review aims to discuss the most recent, conflicting findings regarding the role of intestinal protozoa in gut homeostasis, interactions between intestinal protozoa and the bacterial microbiota, as well as potential immunological consequences of protozoa colonization.
PubMed: 36552252
DOI: 10.3390/biology11121742 -
International Journal of Molecular... Dec 2022is a cosmopolitan intestinal protist colonizing the human gut with varying prevalence depending on the cohort studied and the diagnostic methods used. Its role in human...
is a cosmopolitan intestinal protist colonizing the human gut with varying prevalence depending on the cohort studied and the diagnostic methods used. Its role in human health remains unclear mainly due to the very sporadic number of cross-sectional studies in gut-healthy populations. The main objective of this study was to expand knowledge of the epidemiology of in gut-healthy humans and their animals. A total of 296 stool samples from humans and 135 samples from 18 animal species were analyzed. Using qPCR, a prevalence of 24% was found in humans in contrast to conventional PCR (7%). In humans, several factors were found to influence the prevalence of . A more frequent occurrence of was associated with living in a village, traveling outside Europe and contact with farm animals. In addition, co-infection with spp. was observed in nearly half of the colonized humans. In animals, was detected in 13% of samples from eight species using qPCR. Our molecular phylogenies demonstrate a more frequent occurrence of Genotype 1 in gut-healthy humans and also revealed a likely a new protist species/lineage in rabbits related to and other related organisms.
Topics: Animals; Humans; Rabbits; Cross-Sectional Studies; Dientamoebiasis; Feces; Dientamoeba; Prevalence
PubMed: 36499734
DOI: 10.3390/ijms232315407 -
Revista Espanola de Quimioterapia :... Feb 2023Microscopic examination of the intestinal parasites, from the patient's concentrated feces, has a lower sensitivity when compared to molecular diagnostic techniques....
OBJECTIVE
Microscopic examination of the intestinal parasites, from the patient's concentrated feces, has a lower sensitivity when compared to molecular diagnostic techniques. Therefore, the objective of this study has been to compare both techniques, as well as to evaluate whether there is a correlation between the microscopic examination and the threshold cycles (Ct) obtained for Blastocystis hominis.
METHODS
Retrospective study of the samples received in the Microbiology laboratory during September 2021. The MiniParasep SF® concentration test was performed for microscopic visualization and then PCR was performed with the Seegene AllplexTM Parasite Assay panel.
RESULTS
A 27% (n=74) of the samples were positive by molecular diagnosis, with a total of 87 parasites detected. 53% (n=39) were women with a mean age of 47 ± 24 years. In 76% (n=56) of the cases the service of origin was Primary Care. The most frequently found parasite was B. hominis, 85% (n=64), followed by Dientamoeba fragilis 20% (n=15) and Giardia lamblia 11% (n=8). Co-infection by two parasites was detected in 13 cases (B. hominis + D. fragilis in 6 cases, and B. hominis + G. lamblia in 7 cases). In the microscopic diagnosis, 9.5% (n=26) positivity was obtained. The most frequently found parasite was B. hominis, 84% (n=23), followed by G. lamblia, which was seen in three cases by microscopy. D. fragilis was not seen in any case. Coinfection of B. hominis + G. lamblia was observed in one sample.
CONCLUSIONS
Techniques for molecular diagnosis of intestinal parasites are fast, reliable and more sensitive than microscopic techniques, improving microbiological diagnosis and quality of care.
Topics: Humans; Female; Young Adult; Adult; Middle Aged; Aged; Male; Retrospective Studies; Microscopy; Intestinal Diseases, Parasitic; Giardia lamblia; Feces; Molecular Biology
PubMed: 36453171
DOI: 10.37201/req/088.2022