US 12,007,392 B2
Method and system for detection of mycobacteria in human samples
Irene Grant, Belfast (GB); Antonio Foddai, Abbington, PA (US); John Todd Kuenstner, Abbington, PA (US); Raghava Potula, Philadelphia, PA (US); Tim Bull, Tooting (GB); Saleh Naser, Orlando, FL (US); and Ira Shafran, Winter Park, FL (US)
Assigned to Temple University—Of The Commonwealth System of Higher Education, Philadelphia, PA (US)
Filed by Temple University—Of The Commonwealth System of Higher Education, Philadelphia, PA (US); Queen's University Belfast, Belfast (GB); St. George's, University of London, Tooting (GB); and University of Central Florida, Orlando, FL (US)
Filed on Sep. 27, 2019, as Appl. No. 16/585,391.
Claims priority of provisional application 62/737,609, filed on Sep. 27, 2018.
Prior Publication US 2020/0103403 A1, Apr. 2, 2020
Int. Cl. G01N 33/569 (2006.01)
CPC G01N 33/5695 (2013.01) [G01N 2800/065 (2013.01); G01N 2800/56 (2013.01)] 10 Claims
 
1. A method of treating Crohn's Disease in a subject in need thereof comprising the steps of:
a) obtaining a human blood or peripheral blood mononuclear cell (PBMC) sample from a subject diagnosed with Crohn's Disease;
b) lysing cells of the sample to release any viable target mycobacterial cell;
c) admixing a broad host range mycobacteriophage with the sample under conditions suitable to allow the bacteriophage to infect any viable target mycobacterial cell present in the sample;
d) removing any bacteriophage not infecting a viable target mycobacterial cell;
e) admixing at least a portion of the admixture of step c) following the removal of step d) with bacteria capable of supporting replication of the bacteriophage;
f) plating at least a portion of the admixture of step e) on a substrate to support growth of the bacteria;
g) quantifying the number of bacteriophage plaques that form;
h) discriminating between infected and asymptomatic subjects, comprising identifying the subject as infected when the number of bacteriophage plaques is statistically significantly greater than a reference number of bacteriophage plaques, or as asymptomatic when the number of bacteriophage plaques is not statistically significantly greater than a reference number of bacteriophage plaques, wherein the reference number of bacteriophage plaques is a reference number of plaques in asymptomatic human hosts; and
i) administering a pharmaceutical composition for inhibiting or disrupting microorganism growth or treating a microbial infection to a subject identified as infected.