US 11,788,155 B1
Lung microbiome isolation and cultivation
Andreas Seyfang, Tampa, FL (US); and Nirmal S. Sharma, Tampa, FL (US)
Assigned to UNIVERSITY OF SOUTH FLORIDA, Tampa, FL (US)
Filed by University of South Florida, Tampa, FL (US)
Filed on Jun. 5, 2020, as Appl. No. 16/893,579.
Claims priority of provisional application 62/857,939, filed on Jun. 6, 2019.
Int. Cl. C12Q 1/689 (2018.01); A61K 35/747 (2015.01)
CPC C12Q 1/689 (2013.01) [A61K 35/747 (2013.01); C12Q 2600/106 (2013.01); C12Q 2600/16 (2013.01)] 20 Claims
 
1. A method for preparing an amplified lung microbiome from a subject, comprising
(a) obtaining a bronchoalveolar lavage (BAL) sample from the subject;
(b) culturing a first portion of the BAL sample in a liquid Tryptic Soy broth medium under conditions suitable to expand a first aerobic lung bacteria population;
(c) culturing a second portion of the BAL sample in a liquid Lactobacilli-adapted MRS broth medium under conditions suitable to expand a second aerobic lung bacteria population; and
(d) culturing a third portion of the BAL sample in a modified liquid Chopped Meat medium supplemented with DHNA (1,4-dihydroxy-2-naphthoic acid) under conditions suitable to expand a first anaerobic lung bacteria population,
(e) mixing the first aerobic lung bacteria population, second aerobic lung bacteria population, and the first anaerobic lung bacteria population of steps (b)-(d) produces the amplified lung microbiome, wherein the amplified lung microbiome comprises bacteria from two or more of the following bacterial classes: gammaproteobacteria, fusobacteriia, actinobacteria, bacteroidia, and deltaproteobacteria.