Methods and compositions for detecting infections

A method for detecting whether a subject has an infection comprising:

• (a) incubating a test sample from a subject suspected of having an infection with a labeled nucleoside analog that is preferentially phosphorylated by a thymidine kinase from a pathogenic microorganism for a period of time sufficient to phosphorylate the labeled nucleoside analog by the thymidine kinase of the pathogenic microorganism;
• (b) removing any unphosphorylated or unsequestered labeled nucleoside analog from the test sample;
• and
• (c) detecting labeled nucleoside analog sequestered in the pathogenic microorganism, if any, in the test sample, wherein the presence of labeled nucleoside analog sequestered in the pathogenic microorganism indicates that the subject has an infection.

A kit for detecting the presence of a pathogenic microorganism in a test sample, the kit comprising:

• (a) a labeled nucleoside analog that is preferentially phosphorylated by a thymidine kinase of a pathogenic microorganism;
• (b) a container for contacting the labeled nucleoside analog with a test sample suspected of containing the pathogenic microorganism;
• and
• (c) a device for capturing the pathogenic microorganism, if any, after contacting the labeled nucleoside analog with the sample in the container.

A method for determining whether a subject is a candidate for antibiotic treatment comprising:

• (a) incubating a test sample from a subject suspected of having an infection with a labeled nucleoside analog that is preferentially phosphorylated by a thymidine kinase from a pathogenic microorganism;
• (b) removing any unphosphorylated or unsequestered labeled nucleoside analog from the test sample;
• and
• (c) detecting, in the test sample, labeled nucleoside analog sequestered in the pathogenic microorganism, if any, wherein the presence of labeled nucleoside analog sequestered in the pathogenic microorganism indicates that the subject has an infection and is a candidate for antibiotic treatment.

A method for monitoring the efficacy of an antibiotic treatment in a subject in need thereof comprising:

• (a) incubating a test sample from a subject receiving antibiotic treatment with a labeled nucleoside analog that is preferentially phosphorylated by a thymidine kinase of a pathogenic microorganism;
• (b) removing any unphosphorylated or unsequestered labeled nucleoside analog from the test sample;
• and
• (c) detecting, in the test sample, labeled nucleoside analog sequestered in the pathogenic microorganism, if any, wherein the presence and intensity, if any, of incorporated labeled nucleoside analog sequestered in the pathogenic microorganism provides an indication of the efficacy of the antibiotic treatment.

A method for detecting bacterial or herpes simplex virus (HSV) meningitis in a subject suspected of having such a disease comprising:

• (a) incubating a sample of cerebro-spinal fluid from the subject with 14C-labeled FIAU that is preferentially phosphorylated by a thymidine kinase of a pathogenic microorganism;
• (b) removing any unphosphorylated or unsequestered 14C-labeled FIAU from the sample of cerebro-spinal fluid;
• and
• (c) detecting, in the cerebro-spinal fluid, 14C-labeled FIAU sequestered in the pathogenic microorganism, if any, wherein the presence of incorporated 14C-labeled FIAU sequestered in the pathogenic microorganism indicates that the subject has bacterial or HSV meningitis.

A kit for detecting bacterial or HSV meningitis in a subject suspected of having such a disease, the kit comprising:

• (a) 14C-labeled FIAU;
• (b) a container for contacting the 14C-labeled FIAU with a sample of cerebro-spinal fluid from the subject;
• and
• (c) a device for capturing a pathogenic microorganism containing 14C-labeled FIAU from the sample, if any.

A method for detecting bacterial pneumonia in a subject suspected of having such a disease comprising:

• (a) incubating a sputum sample from the subject with 14C-labeled FIAU that is preferentially phosphorylated by a thymidine kinase of a pathogenic microorganism;
• (b) removing any unphosphorylated or unsequestered 14C-labeled FIAU from the sputum sample;
• and
• (c) detecting, in the sputum sample, 14C-labeled FIAU sequestered in the pathogenic microorganism, if any, wherein the presence of 14C-labeled FIAU sequestered in the pathogenic microorganism indicates that the subject has bacterial pneumonia.

A kit for detecting bacterial pneumonia in a subject suspected of having such a disease, the kit comprising:

• (a) 14C-labeled FIAU;
• (b) a container for contacting the 14C-labeled FIAU with a sputum sample from the subject;
• and
• (c) a device for capturing a pathogenic microorganism containing 14C-labeled FIAU from the sample, if any.

A method for detecting whether a subject has a bacterial infection and, if so, for determining whether an antibiotic has an effect on a pathogenic microorganism causing the bacterial infection, comprising:

• (a) incubating a test sample from a subject suspected of having a bacterial infection with a labeled nucleoside analog that is preferentially incorporated into a pathogenic microorganism in the presence and in the absence of an antibiotic for a period of time sufficient for the pathogenic microorganism to incorporate the labeled nucleoside analog;
• (b) removing any unincorporated labeled nucleoside analog from the test sample;
• (c) detecting the labeled nucleoside analog within the pathogenic microorganism, if any, in the test sample, wherein the presence of labeled nucleoside analog within the pathogenic microorganism indicates that the subject has an infection;
• and
• (d) comparing the amount of labeled nucleoside analog in the pathogenic microorganism incubated in the presence and in the absence of the antibiotic, wherein a reduction in the amount of labeled nucleoside analog in the pathogenic microorganism in the presence of the antibiotic indicates that the antibiotic has an effect on the pathogenic microorganism.

A method for detecting whether a subject has a bacterial infection and, if so, for selecting a treatment for the infection, comprising:

• (a) incubating a test sample from a subject suspected of having a bacterial infection with a labeled nucleoside analog that is preferentially incorporated into a pathogenic microorganism in the presence and in the absence of an antibiotic for a period of time sufficient for the pathogenic microorganism to incorporate the labeled nucleoside analog;
• (b) removing any unincorporated labeled nucleoside analog from the test sample;
• (c) detecting the labeled nucleoside analog within the pathogenic microorganism, if any, in the test sample, wherein the presence of labeled nucleoside analog within the pathogenic microorganism indicates that the subject has an infection;
• and
• (d) comparing the amount of labeled nucleoside analog in the pathogenic microorganism incubated in the presence and in the absence of the antibiotic, wherein a reduction in the amount of labeled nucleoside analog in the pathogenic microorganism in the presence of the antibiotic indicates that the antibiotic is a candidate drug for treating the bacterial infection.

A method for detecting the presence of a pathogenic bacteria in a test sample and, if such a pathogenic bacteria is present in the test sample, for identifying a candidate antibiotic for treating or ameliorating the effects of the pathogenic bacteria, comprising:

• (a) contacting the test sample with a labeled nucleoside analog that is preferentially phosphorylated or incorporated by a thymidine kinase of the pathogenic bacteria;
• (b) further contacting the test sample of step (a) with a panel of antibiotics including one or more controls;
• (c) detecting whether the labeled nucleoside analog has been phosphorylated or incorporated into the pathogenic bacteria;
• and
• (d) comparing the amount of phosphorylated or incorporated labeled nucleoside analog in the pathogenic bacteria exposed to each antibiotic or control in the panel and identifying the antibiotic from the panel, which corresponds to the largest reduction in the amount of phosphorylated or incorporated labeled nucleoside analog in the pathogenic bacteria, wherein the antibiotic identified in step (d) indicates the nature of the bacteria causing the infection and provides an antibiotic selectivity profile to a medical professional.

A kit for detecting the presence of a pathogenic bacteria in a test sample and for determining whether an antibiotic has an effect on the pathogenic bacteria comprising:

• (a) a labeled nucleoside analog that is preferentially phosphorylated by a thymidine kinase of a pathogenic bacteria;
• (b) a panel of antibiotics, including a control;
• (c) a container for contacting the labeled nucleoside analog with a test sample suspected of containing the pathogenic bacteria and the panel of antibiotics, including a control;
• and
• (d) a device for capturing the pathogenic bacteria, if any, after contacting the labeled nucleoside analog with the sample in the container.