Method for evaluating bacterial cell wall integrity
The present invention relates to a method for evaluating the integrity of the cell wall of the bacteria present in a culture in the presence of an antibiotic acting on the bacterial cell wall which, from a practical point of view, allows quickly determining if a bacterium is sensitive or resistant to an antibiotic acting on the bacterial cell wall. Likewise, the present invention also relates to a lysis solution applicable in the preceding method, specifically affecting bacteria having the cell wall damaged by the action of an antibiotic acting on the bacterial cell wall, which allows distinguishing bacteria sensitive to said antibiotic from those resistant to said antibiotic.
1. A method for evaluating sensitivity of a bacterium to an antibiotic acting on the bacterial cell wall which comprises:
i) obtaining a pure culture of the bacterium;
ii) adding to said pure culture of said bacterium an antibiotic;
iii) adding a lysis solution to the culture resulting from step ii), the lysis solution comprising 200 mM 2-amino-2-hydroxymethyl-propane-1,3-diol, a pH of 10, 0.05-3 M of NaCl, and either 0.025% sodium dodecylsulfate or 5% 4-(1,1,3,3-Tetramethylbutyl)phenyl-polyethylene glycol; and
iv) determining the presence of bacterial nucleoid in the culture medium, wherein the presence of bacterial nucleoid in the culture medium is indicative that the bacterium is sensitive to the antibiotic.
2. The method according to claim 1 , wherein the presence of the bacterial nucleoid in the culture medium is carried out by means of staining.
3. The method according to claim 2 , wherein the staining is carried out by means of using one or more fluorochromes.
4. The method according to claim 1 , wherein the antibiotic is selected from the group consisting of a β-lactam antibiotic, an isoniazid, an ethionamide, an ethambutol, a cycloserine and a glycopeptide antibiotic.
5. The method according to claim 1 , wherein the method comprises step v, before or after step (iii), of immobilizing a sample from the culture on a slide.