The cell-wall structure variation in Mycobacterium tuberculosis with different drug sensitivity using Raman spectroscopy in the high-wavenumber region

Abstract

Over the past decade Raman spectroscopy has been successfully used to study the cell structure of various pathogens, including M. tuberculosis (MbT). It is especially important due to the high ability of pathogen to become resistant to anti-tuberculosis drugs usually leading to a lethal outcome. In the present study we investigate deactivated MbT strains with different drug sensitivity (extra-, multi-and sensitive drug resistance) of the Beijing family. Samples have been obtained from patients suffering from pulmonary and extra-pulmonary forms of tuberculosis. Samples have been investigated by Raman spectrometry using a diode-pumped solid-state argon (514 nm) laser excitation source in the high-wavenumber region. As a result, protein and lipid vibrational bands for strain discrimination with various drug resistances have been determined. The obtained results can be used as fast diagnostic tool development for screening MbT strains in clinical medical practice.

Rapid Mycobacterium abscessus antimicrobial susceptibility testing based on antibiotic treatment response mapping via Raman Microspectroscopy

“…Furthermore, the current CAST-R cost per sample for susceptibility testing for two drugs at four different concentrations is approximately $8 (The cost of the required D 2 O is $2, Raman testing is $5, and other experimental materials is $1), a substantially lower cost than that of conventional AST conducted using the broth dilution method ($15 for mycobacterial susceptibility testing of two drugs). Specially, it has been widely reported for the detection of Mycobacterium tuberculosis via Raman Microspectroscopy [ 28 – 30 ]. Thus, the CAST-R may also apply to Mycobacterium tuberculosis which brings greater economic benefits and time savings.…”

Raman spectroscopy for glutathione measurements in Mycobacterium tuberculosis strains with different antibiotic resistance

“…The detailed experimental procedure was described in previous studies. [21,22] The collection of MbT strains consists of strains isolated in the Saint-Petersburg State Research Institute of Phthisiopulmonology. [21,22] For our investigation, we have used strains with different drugs sensitivity obtained from clinical material (sputum and bones for extra-pulmonary strains).…”

“…[21,22] The collection of MbT strains consists of strains isolated in the Saint-Petersburg State Research Institute of Phthisiopulmonology. [21,22] For our investigation, we have used strains with different drugs sensitivity obtained from clinical material (sputum and bones for extra-pulmonary strains). Characteristics of the strains are shown in Table 1, where medium drugresistant (MDR) and extreme drug-resistant (XDR) strains are defined according to WHO classification.…”

“…According methods mentioned in literature, [21,22] all isolates were obtained using a Levenshtein-Jensen medium or Middlebrook 7H9 (BACTEC MGIT 960 automated system) medium. All cultures were harvested at the same growth phases (stationary state).…”

“…Also, the identification of the drug resistance markers that can correlate with bacterial cell death and resistance mechanisms remains a challenge. In our previous investigations, [21,22] we have performed a Raman "screening" for clinical MbT with different drug resistance. One of the promising candidate for such possible markers is GSH revealed in the previous work.…”