Evaluation of Nested Polymerase Chain Reaction and Immunoassay for Rapid Diagnosis of Clostridium difficile in Children with Community Acquired Diarrhea.
BACKGROUND
Clostridium difficile (C. difficile) is a known pathogen associated with diarrhea especially in hospital acquired diarrhea. Yet, it is being recognized as a probable etiology for community acquired diarrhea. The aim of the present study was to detect the presence of C. difficile as a pathogen causing community acquired diarrhea in children and to verify the value of different laboratory methods for diagnosis, namely specific culture, immunoassay for toxin detection, and nested polymerase chain reaction (nested-PCR).


METHODS
This prospective study was carried out in children attending Mansoura University Children's hospital complaining of acute diarrhea between the periods from January 2015 until December 2015. Stool samples were collected and subjected to microbiological culture specific for C. difficile and rapid detection of toxin A by chromatographic device and by nested PCR for toxin B detection.


RESULTS
Stool samples were collected from 413 children. C. difficile was isolated by culture from 41 patients (9.9%), while direct nested-PCR detected 39 (9.4%) toxigenic strains. Toxin A was detected in all 41 positive culture samples (9.9%). In comparing the clinical and laboratory findings between patients with C. difficile and those without C. difficile, though there was a higher association of bloody, watery diarrhea in patients with C. difficile these associations were not statistically significant (p = 0.4, p = 0.5, respectively). Comparing nested-PCR and toxin detection with culture for detection of C. difficile showed an excellent accuracy of both methods.


CONCLUSIONS
From this study, we can conclude that community acquired diarrhea due to C. difficile is common among children. It should be sought among the pathogens causing this infection. Rapid laboratory detection of toxin A by a rapid chromatography device is accurate compared to time consuming culture. Moreover, nested PCR for toxin B is an accurate and rapid method when it is available.
Ahmed
A
Elewa
Maysaa El
ME
Sayed-Zaki
D001426:Bacterial Proteins / Q000235:genetics*; D001427:Bacterial Toxins / Q000235:genetics* / Q000276:immunology*; D002648:Child; D002675:Child, Preschool; D016360:Clostridioides difficile / Q000235:genetics* / Q000276:immunology*; D017714:Community-Acquired Infections / Q000175:diagnosis* / Q000453:epidemiology / Q000382:microbiology; D004269:DNA, Bacterial / Q000235:genetics*; D003967:Diarrhea / Q000175:diagnosis* / Q000453:epidemiology / Q000382:microbiology; D004534:Egypt / Q000453:epidemiology; D004761:Enterocolitis, Pseudomembranous / Q000175:diagnosis* / Q000453:epidemiology / Q000382:microbiology; D004768:Enterotoxins / Q000276:immunology*; D005243:Feces / Q000382:microbiology; D005260:Female; D006776:Hospitals, Pediatric; D006801:Humans; D007118:Immunoassay; D008297:Male; D016133:Polymerase Chain Reaction; D011237:Predictive Value of Tests; D011446:Prospective Studies; D015203:Reproducibility of Results; D057188:Workflow
D001426:Bacterial Proteins; D001427:Bacterial Toxins; D004269:DNA, Bacterial; D004768:Enterotoxins; C041348:tcdA protein, Clostridium difficile; C031815:toxB protein, Clostridium difficile