Systems and processes for distinguishing sequences from microorganisms

• Main Authors: Susan Jisoon Lee, Kyle Rhoden, Sima Mortazavi, Prasanna Thwar Krishnan, Shaokang Zhang, Adam Forrest Allred, Sasan Amini, Ramin Khaksar, Michael Taylor, David Tran, Henrik Gehrmann, Shadi Shokralla, Bryan Thomas Barney
• Format: Patent
• Language: English
• Published: 05.10.2022
• Subjects: BEER; BIOCHEMISTRY; CHEMISTRY; COMPOSITIONS OR TEST PAPERS THEREFOR; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL ORENZYMOLOGICAL PROCESSES; ENZYMOLOGY; MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEICACIDS OR MICROORGANISMS; METALLURGY; MICROBIOLOGY; MUTATION OR GENETIC ENGINEERING; PROCESSES OF PREPARING SUCH COMPOSITIONS; SPIRITS; VINEGAR; WINE

A process for distinguishing infectious from non-infectious microorganisms in a sample comprising receiving a sample comprising a plurality of nucleic acids from at least one microorganism; synthesizing a nucleic acid library from one or more of the nucleic acids; performing amplification on the library with a set of multiplex primers to amplify a plurality of genomic regions of a target microorganism and sequencing the amplicons and determining that the target microorganism in the sample is infectious when amplicons of length greater than 500 base pairs from two or more different regions of a genome of the target microorganism are detected by sequencing and non-infectious when amplicons of greater than 500 base pairs from two or more different regions of the genome are not detected. Also described are methods for establishing the limit of detection of the process using a synthetic SARS-CoV-2 nucleic acid sample and producing amplicons of 100, 300, 500 and 800bp fragments; wherein samples with low viral load (Ct) still show amplicons for all fragments.