Multiplex Compositions And Methods For Quantification Of Human Nuclear DNA And Human Male DNA And Detection of PCR Inhibitors

Abstract

The invention relates to a method for simultaneous quantification of human nuclear DNA and human male DNA in a biological sample while also detecting the presence of PCR inhibitors in a single reaction. The multiplex quantification method also provides a ratio of human nuclear and male DNA present in a biological sample. Such sample characterization is useful for achieving efficient and accurate results in downstream molecular techniques such as genotyping.

We claim: 
     
         1 .- 32 . (canceled) 
     
     
         33 . A method comprising conducting a multiplex real-time Polymerase Chain Reaction (PCR) on a sample, wherein the multiplex is at least a triplex, determining the quantity of human DNA, the level of DNA degradation and the level of PCR inhibitors in the sample; a quantification value, adjusting the amount of sample required to perform a short tandem repeat (STR) genotyping assay based on the quantification value such that an interpretable STR profile is obtained for the majority of the STR loci amplified. 
     
     
         34 . The method of  claim 33 , wherein five (5) amplified STR loci have fragment sizes larger than 140 bases. 
     
     
         35 . The method of  claim 33 , wherein thirteen (13) amplified STR loci have fragment sizes larger than 140 bases. 
     
     
         36 . The method of  claim 33 , wherein the majority of the STR loci amplified are 100-400 bases. 
     
     
         37 . The method of  claim 33 , wherein the majority of the STR loci amplified are 80-250 bases. 
     
     
         38 . The method of  claim 33 , wherein the multiplex is a triplex. 
     
     
         39 . The method of  claim 33 , wherein the sample comprises human male and human female DNA. 
     
     
         40 . The method of  claim 33 , wherein the multiplex real-time PCR of samples from orangutan, macaque, dog, cow, pig, cat, chicken, fish, rabbit, mouse, rat, hamster,  Escherichia coli, Pseudomonas  sp.,  Neisseria gonorrhoeae, Staphyloccus  sp. and  Saccharomyces  sp does not produce amplification products. 
     
     
         41 . The method of  claim 33 , wherein the sample comprises humic acid. 
     
     
         42 . The method of  claim 33 , wherein the sample comprises hematin. 
     
     
         43 . The method of  claim 33 , wherein the sample comprises indigo.