Determining efficiency of a polymerase chain reaction (PCR), useful for clinical diagnosis, comprises estimating the efficiency of PCR in the sample from the dependence of the threshold cycle on the dilution factor for each of the genes

摘要

Determining efficiency of a polymerase chain reaction (PCR), where the number of copies of a nucleic acid sequence in a test sample is determined, comprises amplification of DNA by PCR, registering the number of amplification cycles required to obtain a certain amount of product (CT), and estimating the efficiency of the PCR in the sample from the dependence of CT on the dilution factor. Determining efficiency of a polymerase chain reaction (PCR), where the number of copies of a particular nucleic acid sequence in a test sample is determined, comprises amplification of DNA by PCR of the sample itself, or a diluted stock solution of the sample itself, and one or more controlled dilutions of the sample, and registering the number of amplification cycles required to obtain a certain amount of product (CT), and estimating the efficiency of the PCR in the sample from the dependence of CT on the dilution factor. Independent claims are included for the following: (1) diagnosing and/or classifying a disease by comparing the expression ratio of two genes by determining the ratio of the corresponding mRNAs in a sample; (2) monitoring a diseases progress, where the expressions of two or more genes are compared; (3) making a disease prognosis, where the expressions of two or more genes are compared; (4) comparing the presence of splicing variants of a gene by determining their relative amounts; (5) comparing the activities of alternative promoters by determining the relative amounts of their transcripts; (6) determining the amount of a virus or bacteria in a sample; and (7) diagnostic testing for cancer, including lymphoma, where at lest the kappa:lambda expression is determined