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A multiplex PCR method to detect 14 Escherichia coli serogroups associated with urinary tract infections
1TEDA School of Biological Sciences and Biotechnology, Nankai University, 23 Hongda Street, TEDA, Tianjin 300457, PR China.
Abstract:
Urinary tract infections (UTIs) are one of the most common bacterial infections and are predominantly caused by uropathogenic Escherichia coli (UPEC). E. coli strains belonging to 14 serogroups, including O1, O2, O4, O6, O7, O8, O15, O16, O18, O21, O22, O25, O75 and O83, are the most frequently detected UPEC strains in a diverse range of clinical urine specimens. In the current study, the O-antigen gene clusters of E. coli serogroups O1, O2, O18 and O75 were characterized. A multiplex PCR method based on O-antigen-specific genes was developed for the simultaneous detection of all 14 E. coli serogroups. The multiplex PCR method was shown to be highly specific and reproducible when tested against 186 E. coli and Shigella O-serogroup reference strains, 47 E. coli clinical isolates and 10 strains of other bacterial species. The sensitivity of the multiplex PCR method was analyzed and shown to detect O-antigen-specific genes in samples containing 25 ng of genomic DNA or in mock urine specimens containing 40 colony-forming units (CFUs) per ml. Five urine specimens from hospital were examined using this multiplex PCR method, and the result for one sample was verified by the conventional serotyping methods. The multiplex PCR method developed herein can be used for the detection of relevant E. coli strains from clinical and/or environmental samples, and it is particularly useful for epidemiologic analysis of urine specimens from patients with UTIs.
Insights
A new multiplex PCR method accurately detects 14 common uropathogenic Escherichia coli (UPEC) serogroups. This rapid technique aids in diagnosing urinary tract infections (UTIs) and epidemiological studies.
Area of Science:
- Microbiology
- Molecular Biology
- Infectious Diseases
Background:
- Urinary tract infections (UTIs) are common bacterial infections, frequently caused by uropathogenic Escherichia coli (UPEC).
- UPEC strains from 14 specific O-serogroups are predominantly identified in clinical urine samples.
- Accurate and rapid identification of UPEC serogroups is crucial for UTI diagnosis and management.
Purpose of the Study:
- To characterize O-antigen gene clusters of specific E. coli serogroups (O1, O2, O18, O75).
- To develop and validate a multiplex PCR assay for simultaneous detection of 14 UPEC E. coli serogroups.
- To assess the utility of the multiplex PCR method for clinical and epidemiological analysis of UTIs.
Main Methods:
- Characterization of O-antigen gene clusters for selected E. coli serogroups.
- Development of a multiplex PCR assay targeting O-antigen-specific genes.
- Validation of the multiplex PCR method using reference strains, clinical isolates, and mock urine samples.
Main Results:
- The multiplex PCR method successfully detected all 14 target E. coli serogroups.
- The assay demonstrated high specificity and reproducibility against a panel of bacterial strains and clinical isolates.
- The method showed high sensitivity, detecting as low as 40 CFUs/ml in mock urine specimens.
Conclusions:
- A novel multiplex PCR assay enables the simultaneous detection of 14 clinically relevant E. coli serogroups.
- This method offers a specific, reproducible, and sensitive tool for identifying UPEC strains in clinical and environmental samples.
- The developed multiplex PCR is particularly valuable for epidemiological surveillance of UTIs.
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