Amplifiable metabolic labeling for detection of bacterial viability, growth and antibiotic susceptibility

A method to treat a subject for a bacterial infection comprising:

• a) contacting a sample from a subject suffering from a bacterial infection with an antibiotic;
• b) determining the effect of the antibiotic on incorporation of a single or di-D-amino acid into peptidoglycan of the bacterial cell or trehalose incorporation into the bacterial cell envelope;
• and
• c) administering the antibiotic to said subject, wherein inhibited incorporation of the D-amino acid into peptidoglycan of the bacterial cell or inhibited incorporation of trehalose into the bacterial cell envelope was determined in b), so as to treat said subject for a bacterial infection.

A method to detect antibiotic resistant strains of bacteria in a subject comprising:

• a) contacting a sample from a subject suffering from a bacterial infection with an antibiotic;
• and
• b) determining the effect, if any, of the antibiotic on incorporation of a single- or di-D-amino acid into peptidoglycan of the bacterial cell or trehalose incorporation into the bacterial cell envelope,
• wherein the bacteria are resistant to said antibiotic if said bacteria continue to grow and incorporate the D-amino acid into peptidoglycan of the bacterial cell or trehalose incorporation into the bacterial cell envelope.

A method to detect bacterial contamination of a site comprising,

• a) contacting a sample from a site with an antibiotic;
• and
• b) determining the effect, if any, of the antibiotic on incorporation of a single- or di-D-amino acid into peptidoglycan of the bacterial cell or trehalose incorporation into the bacterial cell envelope,
• wherein an antibiotic that reduces, as compared to control without the antibiotic being present, incorporation of the D-amino acid into peptidoglycan in the bacterial cell or inhibited incorporation of trehalose is indicative of bacterial contamination of said site.

A method to detect low amounts of bacteria in a subject or site comprising:

• a) metabolically labeling bacteria in a sample from a subject or site by incorporation of a single- or di-D-amino acid into peptidoglycan of the bacterial cell, wherein the D-amino acid is labeled with biotin;
• b) capturing said metabolically labeled bacteria with avidin;
• and
• c) detecting the signal with an ELISA, IHC, or western blot,
• wherein the bacteria are grown out/cultured/expanded prior to and/or during metabolic labeling.

A method to detect low amounts of bacteria in a subject or site comprising:

• a) metabolically labeling bacteria in a sample from a subject or site by incorporation of a single- or di-D-amino acid into peptidoglycan of the bacterial cell;
• b) capturing said metabolically labeled bacteria with an antibody;
• and
• c) detecting said antibody,
• wherein the bacteria are grown out/cultured/expanded prior to and/or during metabolic labeling.