Compositions, kits and methods for synthesis and/or detection of nucleic acids

Abstract

A composition comprising a thermostable DNA polymerase; and a PCR inhibitor blocking agent, wherein the PCR inhibitor blocking agent is present in an amount effective to enhance tolerance of an assembled PCR to a PCR inhibitor.

What is claimed is: 
     
         1 . A polymerase chain reaction (PCR) composition, comprising:
 a thermostable DNA polymerase; and   a PCR inhibitor blocking agent,   wherein the PCR inhibitor blocking agent is present in an amount effective to enhance tolerance of an assembled PCR to a PCR inhibitor.   
     
     
         2 . The composition of  claim 1 , wherein the PCR inhibitor blocking agent is a protein. 
     
     
         3 . The composition of  claim 1 , wherein the PCR inhibitor blocking agent is albumin, gelatin, or a combination thereof. 
     
     
         4 . The composition of  claim 1 , wherein the PCR inhibitor blocking agent is bovine serum albumin, fish gelatin, or a combination thereof. 
     
     
         5 . The composition of  claim 1 , wherein up to 50 μM hematin is tolerated in an assembled PCR. 
     
     
         6 . The composition of  claim 1 , wherein up to 15 ng of humic acid per 20 μL reaction volume is tolerated in an assembled PCR. 
     
     
         7 . The composition of  claim 1 , wherein up to 0.01 U/μL heparin is tolerated in an assembled PCR. 
     
     
         8 . The composition of any of the above claims, further comprising a nonionic detergent. 
     
     
         9 . The composition of  claim 8 , wherein the nonionic detergent is selected from TRITON X-100®, Nonidet P-40 (NP-40), Tween 20, and Brij 35. 
     
     
         10 . The composition of  claim 1 , wherein the composition is stable at room temperature for up to 48 hours. 
     
     
         11 . The composition of  claim 10 , wherein the composition is stable at room temperature for up to about 72 hours. 
     
     
         12 . The composition of  claim 1 , wherein the thermostable DNA polymerase is selected from the group consisting of Taq DNA polymerase, Tne DNA polymerase, Tma DNA polymerase, Pfu DNA polymerase, Pwo DNA polymerase, VENT DNA polymerase, DEEPVENT DNA polymerase, mutants or derivatives thereof, and a combination of the foregoing. 
     
     
         13 . The composition of  claim 1 , wherein the concentration of the thermostable DNA polymerase is about 100 to about 500 units per milliliter. 
     
     
         14 . The composition of  claim 1 , further comprising a reagent for hotstart PCR. 
     
     
         15 . The composition of  claim 14 , wherein the reagent is an antibody, an aptamer, a hairpin primer, or a sequestration wax bead. 
     
     
         16 . The composition of  claim 1 , further comprising one or more deoxynucleoside triphosphates. 
     
     
         17 . The composition of  claim 1 , further comprising a passive reference control. 
     
     
         18 . A kit comprising the composition of  claim 1 , further comprising a control nucleic acid sample, and a primer pair specific for PCR amplification of a DNA target in the control nucleic acid sample. 
     
     
         19 . A method for nucleic acid synthesis comprising:
 adding the composition of  claim 1  to a reaction vessel;   adding a nucleic acid sample and a primer to the reaction vessel; and   synthesizing a nucleic acid using the nucleic acid sample as a template.   
     
     
         20 . A method for blocking inhibition of a polymerase chain reaction (PCR) by PCR inhibitors, comprising:
 adding the composition of  claim 1  to a reaction vessel, wherein the composition blocks inhibition of PCR by PCR inhibitors;   adding a nucleic acid sample and a primer to the reaction vessel; and   performing PCR on the nucleic acid sample.   
     
     
         21 . A method for decreasing the run time of a polymerase chain reaction (PCR) by PCR inhibitors, comprising:
 adding the composition of  claim 1  to a reaction vessel, wherein the composition decreases the run time of a PCR;   adding a nucleic acid sample and a primer to the reaction vessel; and   performing PCR on the nucleic acid sample.   
     
     
         22 . The method of  claim 21 , wherein the PCR is a simplex PCR. 
     
     
         23 . The method of  claim 21 , wherein the PCR is a multiplex PCR.