An accurate multiplex antibiotic susceptibility test using a high-resolution CE-SSCP-based stuffer-free multiplex ligation-dependent probe amplification system

Abstract

The success of antimicrobial therapy depends on effective prescription of antibiotics. Assessment of clinical isolates using rapid antimicrobial susceptibility tests allows effective microbiological therapy to be commenced in a timely manner. However, conventional antimicrobial susceptibility testing is time-consuming and laborious. In the present study, we employed stuffer-free multiplex ligation-dependent probe amplification (MLPA) coupled with analysis of single-strand conformation polymorphisms, via high-resolution CE, to develop a multiplex antibiotic susceptibility test. Using this method, parallel analysis of specific genetic markers was employed to determine minimal inhibitory concentration values. The values derived using the stuffer-free MLPA method agreed with those estimated using a conventional broth dilution method. These findings indicate that the stuffer-free MLPA-based approach is a viable alternative to the conventional method.