Methods for Treating and Detecting Sepsis in Humans

Abstract

Biomarkers for identifying sepsis in humans are presented herein, as are related methods, uses, agents, and kits comprising same. Methods for treating, detecting, and diagnosing sepsis in humans are presented herein.

1 . A method comprising
 administering to a human identified as having sepsis a therapeutically effective amount of at least one agent used to treat sepsis,   wherein the human is identified as having sepsis by   analyzing a biological sample isolated from the human for over-representation or under-representation of at least one polynucleotide relative to an internal reference region, wherein the at least one polynucleotide comprises any one of SEQ ID NOs: 1-57 or 94-148 and   wherein the over-representation or under-representation of the at least one polynucleotide in the biological sample is indicative of sepsis, thereby identifying the human as having sepsis.   
     
     
         2 . The method of  claim 1 , wherein the at least one polynucleotide comprising any one of SEQ ID NOs: 1-57 or 94-148 over-represented or under-represented relative to the internal reference region is at least two, at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine, at least ten, at least eleven, at least twelve, at least thirteen, at least fourteen, at least fifteen, at least sixteen, at least seventeen, at least eighteen, at least nineteen, at least twenty, at least twenty-one, at least twenty-two, at least twenty-three, or at least twenty-four of the polynucleotides comprising any one of SEQ ID NOs: 1-57 or 94-148. 
     
     
         3 . The method of  claim 1 , wherein the biological sample is blood, a product derived from blood, or a fraction derived from blood. 
     
     
         4 . The method of  claim 1 , wherein the internal reference region comprises at least one polynucleotide comprising at least one of SEQ ID NOs: 59, 61, or 68. 
     
     
         5 . The method of  claim 1 , wherein detecting the over-representation or under-representation of the at least one polynucleotide relative to an internal reference region comprises at least one of a polymerase chain reaction (PCR)-based detection method, a hybridization-based method, enzyme-linked immunosorbent assay (ELISA), radioimmunoassay (RIA), solid-phase enzyme immunoassay (EIA), mass spectrometry, or microarray analysis. 
     
     
         6 . The method of  claim 5 , wherein the PCR-based detection method is performed using at least one primer pair, wherein each primer pair of the at least one primer pair is specific for any one of SEQ ID NOs: 1-57 or 94-148, and a primer pair specific for at least one of SEQ ID NOs: 59, 61, or 68. 
     
     
         7 . The method of  claim 6 , wherein the primer pair specific for any one of SEQ ID NOs: 1-57 or 94-148 and the primer pair specific for at least one of SEQ ID NOs: 59, 61, or 68 is any one of the primer pairs presented in Tables 1-3. 
     
     
         8 . The method of  claim 7 , further comprising sequencing amplification products corresponding to any one of SEQ ID NOs: 1-57 or 94-148 at least one of SEQ ID NOs: 59, 61, or 68 generated by the PCR-based detection method. 
     
     
         9 . The method of  claim 1 , wherein the at least one polynucleotides comprise circulating nucleic acids. 
     
     
         10 . The method of  claim 1 , wherein the at least one agent used to treat sepsis comprises at least one of an antibiotic, anti-fungal agent, anti-viral agent, anti-parasitic agent, or fluids suitable for intravenous administration. 
     
     
         11 . A primer comprising a manmade nucleotide sequence that binds specifically to a polynucleotide comprising any one of the SEQ ID NOs: listed in Table 4 and at least one manmade tag conjugated thereto, wherein the manmade nucleotide sequence is any one of the polynucleotide primer sequences listed in Tables 1-3 or a variant thereof. 
     
     
         12 . The primer of  claim 11 , wherein the variant of any one of the polynucleotide primer sequences listed in Tables 1-3 is at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, or at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to any one of the polynucleotide sequences listed in Table 4. 
     
     
         13 . The primer of  claim 11 , wherein the manmade tag is a detectable marker. 
     
     
         14 . The primer of  claim 11 , the primer consisting essentially of or consisting of a manmade nucleotide sequence that binds specifically to a polynucleotide of any one of the SEQ ID NOs: listed in Table 4 and at least one manmade tag conjugated thereto, wherein the manmade nucleotide sequence is any one of the polynucleotide primer sequences listed in Tables 1-3 or a variant thereof and at least one manmade tag conjugated thereto, wherein the manmade nucleotide sequence is any one of the polynucleotide primer sequences listed in Tables 1-3 or a variant thereof. 
     
     
         15 . The polynucleotide primer sequence of  claim 14 , wherein the variant of any one of the polynucleotide primer sequences listed in Tables 1-3 is at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, or at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to any one of the polynucleotide primer sequences listed in Tables 1-3. 
     
     
         16 . The primer of  claim 14 , wherein the manmade tag is a detectable marker. 
     
     
         17 . A kit for detecting sepsis in a human comprising at least one primer pair of  claim 11  and instructions for use thereof. 
     
     
         18 . The kit of  claim 17 , wherein the at least one primer pair comprises four primer pairs and wherein each primer pair of the four primer pairs specifically amplifies a different polynucleotide comprising any one of the SEQ ID NOs: listed in Table 4.