Method for diagnosing tuberculosis in a urine sample
A technology for tuberculosis, pulmonary tuberculosis, applied in the field of diagnosing tuberculosis in urine samples, which can solve the problems of inability to distinguish latent TB, unvalidated biomarkers, and no commercially available point-of-care tests for diagnosing TB
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Image
Examples
Embodiment 1
[0061] Example 1: Identification of Mycobacterium tuberculosis-derived proteins that are selectively present in urine samples from patients with active TB, latent TB infection, or non-TB disease
[0062] method
[0063] Material
[0064]Acetonitrile and formic acid were both HPLC analytical grade and were purchased from Thermo Fisher Scientific (Waltham, MA). 18.0 MΩ water was produced using the Mili-Q UV Plus water purification system. Dithiothreitol (DTT), ammonium bicarbonate and iodoacetamide (IAA) were all purchased from Sigma Aldrich (St. Louis, MO, USA). Trypsin was purchased from Promega (Madison, WI, USA).
[0065] Urine Collection and Clinical Case Definitions
[0066] Urine samples were collected from patients seen in South African clinics who were suspected of being infected with TB ( Figure 1a ). Informed consent was obtained from all participants and the study was approved by the Health Sciences Human Research Ethics Committee of the University of Cape T...
Embodiment 2
[0131] Example 2: Identification of Mycobacterium tuberculosis-derived proteomes present in patient urine samples that differentiate active TB, latent TB infection, and non-TB / non-LTBI disease
[0132] The data from Example 1 regarding the frequency at which tryptic peptides derived from individual M.tb proteins were observed in pooled and individual urine samples were collated and presented in Table 7, from which apparently no to one M.tb protein present in every sample from a given group and absent in all other samples (e.g. present in all active TB patient urine samples but not in all LTBI and non-TB / non- -absent in LTBI samples). Of note, the proteins with the highest frequencies observed in the various urine samples showed relatively poor discrimination between the three patient groups. For example, Rv0765c was observed in 4 of 5 TB samples (consisting of 2 of 2 pooled samples and 2 of 3 individual samples), but not in 3 non-TB / non-LTBI samples Also observed in 2 of . ...
Embodiment 3
[0138] Example 3: Targeted identification of proteins derived from Mycobacterium tuberculosis in patient urine samples
[0139] Sample Preparation
[0140] Urine samples were collected from 64 individual TB suspects recruited from 3 patient groups of approximately equal size: as in Example 1, the patient groups were active TB, LTBI and non-TB / non-LTBI; all patients were HIV negative of. Proteins from individual urine samples (10ml per sample) were fractionated first through 15ml filters of 50kDa molecular weight cut-off (MWCO) essentially as described in Example 1, followed by 15ml 3kDa MWCO filters followed by 0.5ml 3kDa MWCO The filtrate was concentrated by a filter. Protein (approximately 30 μg) from each sample was proteolyzed using trypsin by filter assisted sample preparation (FASP) according to literature protocol (37). Tryptic peptides were recovered for each individual sample and desalted prior to LC-MS / MS analysis. Equal amounts of peptides from each of the 64 ...
PUM
| Property | Measurement | Unit |
|---|---|---|
| mass | aaaaa | aaaaa |
| molecular weight | aaaaa | aaaaa |
Abstract
Description
Claims
Application Information
Login to View More