Novel Methods for Rapid Identification and Susceptibility Testing of Bacteria

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Background

As strain variation and drug resistance become more pervasive, the prevention and control of infections has become a serious problem in recent years. Traditional pathogen detection methods, such as analysis and identification of microorganisms based on biochemical and serological characteristics, cannot meet clinical needs.
Rapid detection of pathogenic bacteria is key to the diagnosis process. Modern technologies such as microarray, Raman spectroscopy, mass spectrometry, DNA sequencing as well as multiplex real-time PCR are becoming the most used tools within this field, which will accelerate the development of the health industry.

DNA microarrays offer the potential for simultaneous detection of many pathogenic bacteria on a solid substrate such as glass. Due to the high throughput attainable through miniaturization, these tools are best suited for revealing the presence or absence of genetic characteristics of specific pathogenic bacteria and allow a fast response during an epidemic or outbreak.
The recently developed surface-enhanced Raman scattering (SERS) is a powerful and promising tool in bacterial identification because of its high sensitivity, high speed, relatively low cost, and portability. Several pathogenic species have been identified by SERS, such as Klebsiella pneumoniae, Escherichia coli and Staphylococcus aureus. The growth of media-cultured clinical isolates can be distinguished by their SERS-fingerprinting spectra when combined with multivariate data analysis procedures. The major vibrational bands of the observed bacterial spectra are attributed to the entire structure of the organism, such as their nucleic acids, proteins, lipids, and carbohydrates. However, because drug-resistant and susceptible strains of bacteria have similar components, SERS has been scarcely adopted to distinguish drug-resistant bacterial pathogens.

Mass spectrometry is another revolutionary approach for the analysis of microorganisms. The advantages of this method include easy manipulation, low cost and accurate analysis. Therefore, within just a few years, MS has become the worldwide leading solution for clinical microbiology to identify cultured bacteria. In addition to identification, MS has also been used for bacterial subtyping and identification of antimicrobial resistance.

Sequence-based identification of microorganisms is more objective and accurate than conventional methods, especially for detecting mutation sites of resistance genes and for classifying unusual microorganisms. As DNA sequencing allows to determine the precise order of nucleotides within a bacterial genome, it has greatly accelerated medical research and discovery.
Multiplex real-time PCR combines the advantages of two techniques: multiplex PCR and real-time PCR, which can be used for the simultaneous and rapid screening of microorganisms. The application of this method to clinical samples increases the sensitivity for disease diagnosis on direct clinical material and infection control.

This Research Topic aims to cover new applications of these technologies for rapid, high-throughput detection of pathogenic bacteria and drug resistance detection.


Keywords: SERS, microarray, MS, bacteria identification, susceptible testing

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