A simplified multiplex PCR-based typing method for common Salmonella enterica serovars supported by online...

Area of Science:

• Microbiology
• Molecular Biology
• Bioinformatics

Background:

• Traditional Salmonella serotyping is laborious and time-consuming.
• A need exists for a faster, simpler alternative method for Salmonella serovar identification.

Purpose of the Study:

• To improve and simplify a multiplex PCR-based genome typing (MPGT) method for Salmonella.
• To develop an online server for rapid Salmonella serovar determination using MPGT.

Main Methods:

• Standardized a multiplex PCR-based genome typing (MPGT) method using 59 Salmonella isolates of 31 serovars.
• Modified existing primers for improved accuracy and separated reactions for electrophoresis.
• Validated the method through a blind study with 81 isolates and confirmed with whole genome data from 440 isolates.
• Established a public online server for data storage and comparison of MPGT patterns.

Main Results:

• Identified 16 target genes with variable presence across serotypes, enabling unique amplification patterns for genetically related strains.
• Developed a multiplex PCR assay that generates specific binary codes for 31 Salmonella serovars.
• Predicted 83 different MPGT codes in silico based on whole genome information of 440 strains.
• Confirmed that major serovars typically exhibit unique MPGT codes.

Conclusions:

• The multiplex PCR assay provides specific binary codes for Salmonella serovars.
• The online server facilitates comparison of obtained PCR results with stored patterns.
• The MPGT method is simple, rapid, and cost-effective, making it valuable for outbreak management.