[Detection and identification of seven clinical common pathogenic bacteria by oligonucleotide microarray]

Area of Science:

• Microbiology
• Molecular Biology
• Diagnostics

Background:

• Accurate and rapid detection of clinical pathogenic bacteria is crucial for effective treatment and public health.
• Existing diagnostic methods can be time-consuming and labor-intensive.

Purpose of the Study:

• To develop and validate an oligonucleotide microarray system for the simultaneous detection of seven common clinical pathogenic bacteria.
• To assess the performance of the microarray in clinical samples.

Main Methods:

• Utilized double polymerase chain reaction (PCR) to amplify 16S and 23S ribosomal DNA (rDNA) gene segments.
• Constructed an oligonucleotide microarray for simultaneous detection of specific pathogens: EHEC O157:H7, Vibrio parahaemolyticus, Salmonella sp., Vibrio cholerae, Listeria monocytogenes, Campylobacter jejuni, and Shigella sp.
• Evaluated specificity, sensitivity, and reproducibility of the microarray, and tested it on 81 patient stool specimens.

Main Results:

• The double PCR method successfully amplified target 16S and 23S rDNA sequences for all seven pathogens.
• The oligonucleotide microarray demonstrated a sensitivity of 10^3 CFU/ml with high specificity, showing no cross-reactivity with non-targeted bacteria.
• The assay achieved a positive detection rate of 39.5% in stool specimens, with high coincidence rates (96.3% overall, 96.8% for identification) compared to routine bacteriological examinations.

Conclusions:

• The established oligonucleotide microarray assay provides a convenient, rapid, accurate, stable, and high-throughput method for detecting seven key pathogenic bacteria.
• This assay is well-suited for clinical specimen examination and epidemiological field investigations, offering significant advantages over traditional methods.