Quantitative nucleic acid assay
Quantitative nucleic acid assay for a nucleic acid (A) in an individual sample material (PP) comprises: (1) adding a known amount of \- 1 standard nucleic acid (S), with a different nucleotide sequence to A, to the sample material PP; (2) if necessary, converting S into a target nucleic acid ST and...
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Zusammenfassung: | Quantitative nucleic acid assay for a nucleic acid (A) in an individual sample material (PP) comprises: (1) adding a known amount of \- 1 standard nucleic acid (S), with a different nucleotide sequence to A, to the sample material PP; (2) if necessary, converting S into a target nucleic acid ST and converting A into a target nucleic acid AT; (3) generating multiple copies XS from S or ST and multiple copies XA from A or AT; (4) determining a measured value MPP(XA)m for the amount of XA and a measured value MPP(XS)m for the amount of XS; (5) preparing n standards PR(n), where n = \- 1 and each standard comprises a defined amount of \- 1 of the standards S but is not derived from the sample material PP; (6) treating PR(n) under the reaction conditions of steps (2)-(4), in several batches or in a single batch when n > 1, to obtain \- 1 measured value MPR(XS)m for each standard; (7) establishing a calibration curve E on the basis of the n measured values MPR(XS)m; (8) comparing MPP(XS)m with the value corresponding to the same amount on the calibration curve to obtain a correction factor f, and either (a) correcting MPP(XA)m by the correction factor f to obtain a corrected value MPP(XA), and determining the amount of A by comparing MPP(XA) with the calibration curve, or (b) establishing a corrected calibration curve E' using the correction factor and determining the amount of A by comparing MPP(XA)m with the corrected calibration curve. |
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