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Simultaneous Molecular Detection of Common Bacterial Enteropathogens in Children with Diarrhea by Multiplex-PCR Assay
Insights
A new multiplex PCR assay effectively detects common bacterial causes of childhood diarrhea, including Shigella and Campylobacter jejuni. This rapid molecular method offers high sensitivity and specificity for identifying enteropathogens in fecal samples.
Area of Science:
- Microbiology
- Molecular Diagnostics
- Pediatric Infectious Diseases
Background:
- Diarrheal diseases are a leading cause of mortality in children globally, particularly in low- and middle-income countries.
- Accurate and rapid identification of bacterial enteropathogens is crucial for effective treatment and control.
Purpose of the Study:
- To develop and validate a multiplex PCR assay for the simultaneous detection of key bacterial enteropathogens in pediatric fecal specimens.
- To evaluate the assay's sensitivity and specificity for common gastrointestinal pathogens.
Main Methods:
- A multiplex PCR assay was designed to detect Shigella spp., Campylobacter jejuni, Enteropathogenic Escherichia coli (EPEC), Enterotoxigenic Escherichia coli (ETEC), and Salmonella enterica.
- One hundred and three stool samples from children under five with gastroenteritis were analyzed.
Main Results:
- The assay detected Shigella spp. (21.35%), Campylobacter jejuni (10.67%), EPEC (1.94%), and ETEC (0.97%). Salmonella enterica was not detected.
- Coinfections, such as Shigella spp. with Campylobacter jejuni and EPEC with Campylobacter jejuni, were identified.
- The multiplex PCR demonstrated high analytical sensitivity (0.01 ng/µL DNA) and 100% analytical specificity.
Conclusions:
- The developed multiplex PCR assay is a rapid, sensitive, and specific tool for detecting common bacterial enteropathogens.
- This molecular method can aid in the timely diagnosis of infectious diarrhea in children.
Background:
Diarrhea remains a major threat to children in low- and middle-income countries, which is the second cause of death among children in the world. The aim of the present study was to develop and evaluate a multiplex-PCR assay for direct detection of common bacterial enteropathogens in fecal specimens.
Methods:
One hundred and three stool specimens were collected from children under 5 years of age with gastroenteritis during a six-month period in Ilam, Iran. The multiplex PCR assay simultaneously detected Shigella spp., Campylobacter jejuni, Enteropathogenic Escherichia coli (EPEC), Enterotoxigenic Escherichia coli (ETEC), and Salmonella enterica in stool samples.
Results:
Our results demonstrated that the prevalence of Shigella spp. Campylobacter jejuni, EPEC, ETEC, and Salmonella enterica were 21.35%, 10.67%, 1.94%, 0.97% and 0%, respectively. In addition, Shigella spp. with Campylobacter jejuni and EPEC with Campylobacter jejuni coinfection were observed in sample 11 (10.67%). The analytical sensitivity of the multiplex PCR assay was estimated to be 0.01 ng/µL of genomic DNA from culture. The analytical specificity was determined to be 100% by using common and standard enteropathogenic bacterial strains.
Conclusions:
The molecular method developed in the study was rapid, sensitive, and specific for detection of common bacterial enteropathogens.
