Control nucleic acids for multiple parameters

Abstract

The present invention concerns the amplification of at least a first and a second target nucleic acid that may be present in at least one fluid sample using an internal control nucleic acid for qualitative and/or quantitative purposes.

What is claimed is: 
     
         1 . A process for isolating and simultaneously amplifying at least a first and a second target nucleic acid that may be present in one or more samples, the process comprising the steps of:
 a) adding an internal control nucleic acid to each of the one or more samples;   b) combining a solid support material and the one or more samples in one or more vessels for a period of time, under conditions sufficient to permit nucleic acids comprising the target nucleic acids and the internal control nucleic acid to be immobilized on the solid support material;   c) isolating the solid support material from the other material present in the samples in a separation station;   d) purifying the target nucleic acids in the separation station and washing the solid support material one or more times with a wash buffer;   e) contacting the purified target nucleic acids and the purified internal control nucleic acid with one or more amplification reagents, comprising at least one distinct set of primers for each of the target nucleic acids and for the internal control nucleic acid in at least two reaction vessels, wherein at least a first reaction vessel comprises at least the first target nucleic acid, and at least a second reaction vessel comprises at least the second target nucleic acid, and wherein the second target nucleic acid is absent from the first reaction vessel;   f) incubating in the reaction vessels the purified target nucleic acids and the purified internal control nucleic acid with the one or more amplification reagents for a period of time, under conditions sufficient for an amplification reaction indicative of the presence or absence of the target nucleic acids to occur; and   g) detecting and measuring signals generated by the amplification products of the target nucleic acids and being proportional to the concentration of the target nucleic acids, and detecting and measuring a signal generated by the internal control nucleic acid; wherein the conditions for amplification and detection in steps d) to g) are identical for the at least first and second purified target nucleic acids and the internal control nucleic acid, and wherein the sequence of the internal control nucleic acid is identical for the at least first and second purified target nucleic acids.   
     
     
         2 . The process according to  claim 1 , wherein the presence of an amplification product of the internal control nucleic acid is indicative of an amplification occurring in the reaction mixture even in the absence of amplification products for one or more of the target nucleic acids. 
     
     
         3 . The process according to  claim 1 , further comprising the following step: h) determining the quantity of one or more of the target nucleic acids. 
     
     
         4 . The process according to  claim 1 , wherein the amplification reagents comprise a polymerase with reverse transcriptase activity, the process further comprising between step e) and step f) the step of incubating in the reaction vessels the purified nucleic acids with the one or more amplification reagents for a period of time and under conditions suitable for transcription of RNA by the polymerase with reverse transcriptase activity to occur. 
     
     
         5 . The process according to  claim 1 , wherein the internal control nucleic acid is DNA. 
     
     
         6 . The process according to  claim 1 , wherein the internal control nucleic acid is RNA. 
     
     
         7 . The process according to  claim 1 , wherein the internal control nucleic acid is an armored nucleic acid. 
     
     
         8 . The process according to  claim 1 , wherein the sequence of the internal control nucleic acid is different from the sequences of the other nucleic acids present in the one or more samples. 
     
     
         9 . The process according to  claim 1 , wherein the sequence of the internal control nucleic acid is derived from a naturally occurring genome. 
     
     
         10 . The process according to  claim 1 , wherein the sequence of the internal control nucleic acid is scrambled. 
     
     
         11 . The process according to  claim 1 , wherein the internal control nucleic acid has a melting temperature from 50° C. to 90° C. 
     
     
         12 . The process according to  claim 1 , wherein the internal control nucleic acid has a length of up to 500 bases. 
     
     
         13 . The process according to  claim 1 , wherein the sequence of the internal control nucleic acid has a GC content of 30% to 70%. 
     
     
         14 . The process according to  claim 1 , wherein the internal control nucleic acid has a concentration between LOD and 20×LOD. 
     
     
         15 . The process according to  claim 1 , wherein the internal control nucleic acid has a concentration between 20×LOD and 5000λLOD. 
     
     
         16 . The process according to  claim 1 , wherein more than one internal control nucleic acid is added in step a), but only one of the internal control nucleic acids is amplified in step f).