Stool Of Pediatric Patients Research Articles

Fusobacteriumnucleatum Adheres to Clostridioides difficile via the RadD Adhesin to Enhance Biofilm Formation in Intestinal Mucus

Although Clostridioides difficile infection (CDI) is known to involve the disruption of the gut microbiota, little is understood regarding how mucus-associated microbes interact with C difficile. We hypothesized that select mucus-associated bacteria would promote C difficile colonization and biofilm formation. To create a model of the human intestinal mucus layer and gut microbiota, we used bioreactors inoculated with healthy human feces, treated with clindamycin and infected with C difficile with the addition of human MUC2-coated coverslips. C difficile was found to colonize and form biofilms on MUC2-coated coverslips, and 16S rRNA sequencing showed a unique biofilm profile with substantial cocolonization with Fusobacterium species. Consistent with our bioreactor data, publicly available data sets and patient stool samples showed that a subset of patients with C difficile infection harbored high levels of Fusobacterium species. We observed colocalization of C difficile and F nucleatum in an aggregation assay using adult patients and stool of pediatric patients with inflammatory bowel disease and in tissue sections of patients with CDI. C difficile strains were found to coaggregate with F nucleatum subspecies invitro; an effect that was inhibited by blocking or mutating the adhesin RadD on Fusobacterium and removal of flagella on C difficile. Aggregation was shown to be unique between F nucleatum and C difficile, because other gut commensals did not aggregate with C difficile. Addition of F nucleatum also enhanced C difficile biofilm formation and extracellular polysaccharide production. Collectively, these data show a unique interaction of between pathogenic C difficile and F nucleatum in the intestinal mucus layer.

The incidence of Shigella and Salmonella in the stool of pediatric patients

Objective: Gastroenteritis is a common illness in the pediatric age group, the causes could be viral, bacterial and fungal. This study was focused on bacterial pathogens like Shigella and Salmonella.Methods: About 1399 stool samples were collected in a period between January 2010 and May 2011 at the child welfare hospital, all the samples were cultured on suitable culture media and then biochemical activities were tested using API-E 20 and sugar fermentation tests, the isolates were sent for sensitivity test with a group of antibiotics.Results: Out of 1399 cases 50 cases only showed a positive growth for Shigella and Salmonella as pathological microorganisms.Conclusion: The incidence of Shigella and Salmonella was considered low when compared to other causes of gastroenteritis among children as enteropathogenic Escherichia coli or viral causes.

A Novel Sample Processing Method for Rapid Detection of Tuberculosis in the Stool of Pediatric Patients Using the Xpert MTB/RIF Assay.

BackgroundTuberculosis (TB) is difficult to diagnose in children using molecular tests, because children have difficulty providing respiratory samples. Stool could replace sputum for diagnostic TB testing if adequate sample processing techniques were available.MethodsWe developed a rapid method to process large volumes of stool for downstream testing by the Xpert MTB/RIF (Xpert) TB-detection assay. The method was tested and optimized on stool samples spiked with known numbers of M. tuberculosis colony forming units (CFU), and stools from M. tuberculosis-infected cynomolgus macaques (Macaca fascicularis). Performance was scored on number of positive Xpert tests, the cycle thresholds (Cts) of the Xpert sample-processing control (SPC), and the Cts of the M. tuberculosis-specific rpoB probes. The method was then validated on 20 confirmed TB cases and 20 controls in Durban, South Africa.ResultsThe assay’s analytical limit of detection was 1,000 CFU/g of stool. As much as one gram of spiked stool could be tested without showing increased PCR inhibition. In analytical spiking experiments using human stool, 1g samples provided the best sensitivity compared to smaller amounts of sample. However, in Macaques with TB, 0.6g stool samples performed better than either 0.2g or 1.2g samples. Testing the stool of pediatric TB suspects and controls suggested an assay sensitivity of 85% (95% CI 0.6–0.9) and 84% (95% CI 0.6–0.96) for 0.6g and 1.2g stool samples, respectively, and a specificity of 100% (95% CI 0.77–1) and 94% (95% CI 0.7–0.99), respectively.ConclusionThis novel approach may permit simple and rapid detection of TB using pediatric stool samples.

Gastrointestinal pathogens detected by multiplex nucleic acid amplification testing in stools of pediatric patients and patients returning from the tropics.

Gastrointestinal infections are caused by a broad spectrum of pathogens. Conventional diagnostic procedures are resource and time consuming due to single pathogen testing, often in different laboratories. We analyzed 312 consecutive stool samples from pediatric patients (n = 127) with gastroenteritis or from adult travelers returning from the tropics with suspected parasite infestation (n = 185) using commercial multiplex nucleic acid amplification testing (NAT) (xTAG gastrointestinal pathogen panel, Luminex) covering 15 diarrhea-causing pathogens. The results of the positive samples and a representative number of negative samples were compared to standard methods, including NAT, direct antigen detection (DAD), bacterial culture and microscopy. Of the 185 samples from adult travelers, 21 (11 %) were multiplexNAT-positive, with enterotoxigenic Escherichia coli (4 %) being the predominant pathogen. Microscopic examination revealed Blastocystis hominis in 23 % not covered by the panel. MultiplexNAT scored positive in 66 pediatric samples (52 %), with rotavirus (27 %) being the most prevalent. All adenovirus-, rotavirus-, Clostridium difficile- and Cryptosporidium-positive samples were confirmed in external laboratories, but only 40 % of norovirus- and 29 % of Giardia-positive samples. Analysis of frozen specimens by bacterial culture showed the highest discrepancies with the multiplexNAT. Our study demonstrates broad detection of relevant gastroenteritis pathogens by multiplexNAT with a short turnaround time. This is important for diagnosis, infection control and empiric management of gastroenteritis patients, but may be selectively complemented by bacterial culture and resistance testing.

Percentage of Rotavirus infection in stool of pediatric patients at Children Welfare Teaching Hospital in Baghdad

Background: Gastroenteritis is a common illness in pediatric age group; the causes could be bacterial, fungal or viral infection. Rotavirus is the most common cause of infectious diarrhea in children. Children between 3 months to 2 years old have the most severe symptoms. Rotavirus is extremely easy to catch and is transmitted mainly by the oral- fecal route. For diagnosis; the specimen of choice is the stool which has been collected during the first 3 to 5 days of illness. If collected 8 or more days after onset of symptoms the specimen will rarely contain the virus.
 Objective: The objective of this study includes a survey for Rotavirus that cause diarrhea in children admitted to Children Welfare Teaching Hospital in Baghdad between 13th. of Juanuary-2010 to 14th.of October-2010 using a rapid test method for detection of Rota virus to children from one day to 6 years.
 Materials and Methods: Latex agglutination test for the stool of 61 children suspected with Rotavirus infection was done in addition to general and bacteriological test of these stool samples at Children Welfare Teaching Hospital from January 2010 to October 2010
 Results: There were 61 children from one day-6 years old, only 11(18.03%) had positive Rotavirus infection with high percentage in males (seven cases) and (four cases) in females and only one case detected with positive Rotavirus infection at the age of 42 month. No pathogenic bacteria or fungus were isolated.
 Conclusion: The data indicate that male has highly incidence than female and children up to 78 months had diarrhea that not caused by Rotavirus.

Percentage of Rotavirus infection in stool of pediatric patients at Children Welfare Teaching Hospital in Baghdad

Percentage of Rotavirus infection in stool of pediatric patients at Children Welfare Teaching Hospital in Baghdad

Significance of Seasonal Influenza Viruses in the Stool of Pediatric Patients

Significance of Seasonal Influenza Viruses in the Stool of Pediatric Patients

Polymerase chain reaction for Cryptosporidium in the immunocompromised host

Sirs,I read with great interest the article by Bandin et al. entitled“Cryptosporidiosis in paediatric renal transplantation” [1]. Iam grateful for their contribution to the knowledge ofCryptosporidium’s impact on pediatric kidney transplanta-tion. However, I would like to make some comments.First, the diagnosis in the immunocompromised host ismore difficult than in the immunocompetent one. SpecificZiehl–Nielsen staining techniques have low sensitivity, espe-cially comparedwith polymerase chain reaction[2]. So manycases in kidney transplantation can be missed; therefore, thereal prevalence in this cohort could have been higher. Thisshould alert everyone to rule out with appropriate diagnostictesting the presence of Cryptosporidium parvum in the stoolsof pediatric patients who receive kidney transplantation.Second, no drug has proven efficacy in the severelyimmunocompromised host against C. parvum. Unlikelypositive results can be achieved with nitazoxanide [3],paromomycin [4] or the combination of paromomycin andazithromycin [5]. The best efficacy is obtained by loweringimmunosuppression [6].References

Molecular detection of the entire spectrum of human adenoviruses by a two-reaction RQ-PCR assay

Adenoviruses (AdV) can cause life-threatening infections in immunosuppressed patients, particularly in the allogeneic stem cell transplantation (SCT) setting. Reliable diagnostic tests, covering the entire spectrum of the currently known 51 human AdV serotypes, are therefore of paramount importance. We have recently published a five-reaction real-time (RQ-) PCR assay detecting the AdV species A-F, and have demonstrated its ability to predict disseminated AdV disease. However, for routine diagnostic purposes it was desirable to develop a more economic approach to reducing cost and labor. We have sequenced presumably conserved regions of the adenoviral genome, and developed a two-reaction RQ-PCR assay. Due to the genetic similarities between the species B, D and E we could establish an assay facilitating detection and quantification of these three species. Similarly, a second assay covering the species A, C and F, has been established. The detection systems were tested using reference strains from each serotype and clinical samples derived from peripheral blood and stool of pediatric patients after SCT. The specificity and sensitivity of the two-reaction assay is comparable to the published five-reaction RQ-PCR assay, and permits quantification of AdV-DNA down to 1fg per reaction. From the perspective of routine clinical diagnosis, the assay provides an improvement by facilitating early detection and monitoring of adenoviral infections at a substantially reduced cost.