Patent ID: 6331417
Filing Date: 2001-12-18
Classification: A61P,C12Q

Abstract:
A method of determining the amount of a target nucleic acid having the sequence shown in SEQ ID NO:1 in a sample which method comprises:(i) mixing the sample with a predetermined amount of control nucleic acid;(ii) bringing the mixture formed in (i) into contact with a pair of nucleic acid molecules each comprising:a primer portion consisting of a contiguous sequence of from 10 to 50 nucleotides that hybridizes to (a) the target nucleic acid molecule represented by SEQ ID NO:1, or (b) to the complement of the molecule of SEQ ID NO:1 in a polymerase chain reaction (PCR) buffer comprising 16 mM (NH.sub.4).sub.2 SO.sub.4, 67 mM Tris-HCl (pH 8.8 at 25.degree. C.), 0.01% Tween-20, 2 mM MgSO.sub.4 at 50.degree. C.; and optionally a further portion comprising from 1 to 25 nucleotides joined to and immediately 5' to the 5' end of the primer portionwherein one nucleic acid molecule has a primer portion of type (a) and the other nucleic acid molecule has a primer portion of type (b); and wherein the two primers in combination amplify the target nucleic acid molecule represented by SEQ ID NO:1, or a section thereof, in a polymerase chain reaction;(iii) performing a nucleic acid amplification reaction, said reaction requiring the presence of the primer portions defined in (ii) to amplify the target nucleic acid sequence or a section thereof and the control nucleic acid or a section thereof;(iv) determining the relative quantities of the amplified control and target nucleic acids; and(v) calculating from the determination of (iv) the amount of target nucleic acid in the sample.