Patent ID: 11965216
Assignee: POLYSKOPE LABS
Field: Biotechnology (Chemistry)
Classification: CPC C | IPC C

Claim 0:
1. A method comprising:
a. enriching a food sample comprising a plurality of pathogens by co-culturing the plurality of pathogens together in a rich and nonselective media to form an enriched sample; wherein the rich and nonselective media comprises components to promote the growth of the plurality of pathogens; wherein the plurality of pathogens comprises at least two pathogens, wherein at least one of the plurality of pathogens is a member of the group consisting of Listeria monocytogenes, E. coli, and Salmonella spp.;
b. conducting a first sample lysis and a second sample lysis on the enriched sample or a portion thereof, wherein the second sample lysis is performed at a temperature higher than a temperature of the first sample lysis, thereby forming an unpurified lysed sample comprising the DNA of at least one of Listeria monocytogenes, E. coli, and Salmonella spp.;
c. conducting a multiplex amplification with amplification primers on at least a portion of the unpurified lysed sample, wherein the amplification primers comprise at least three primer pairs, wherein a first primer pair is complementary to a target nucleic acid sequence of Listeria monocytogenes, wherein a second primer pair is complementary to a target nucleic acid sequence of E. coli, wherein a third primer pair is complementary to a target nucleic acid sequence of Salmonella spp., and wherein the multiplex amplification is conducted in one tube; and
d. detecting a presence or absence of each of Listeria monocytogenes, E. coli, and Salmonella spp. from a result of the multiplex amplification in the one tube; and wherein a, b, c, and d are performed in a positive total time of about 28 hrs or less;
wherein the rich and nonselective media comprises per 1 L of water at least one of:
(a) yeast extract at a concentration between about 12 g/L and about 20 g/L,
(b) pancreatic digest of casein at a concentration between about 34 g/L and about 50 g/L, or
(c) enzymatic digest of soy at a concentration between about 6 g/L and about 10 g/L.