| 摘要 |
596252 Disclosed is a method for detecting and/or typing and/or quantitation simultaneously the presence of HPV 18, 52, 59, 39, 51, 56, 66, 16, 45, 58, 31, 33 and 35 in a biological sample, the method comprising the following four parallel reactions: Reaction 1: a) amplification of the nucleic acid fragments of HPV 18, 52 and 59 in the presence of: (i) a nucleic acid polymerase (ii) a forward PCR primer with an oligonucleotide sequence of SEQ ID NO:1, (iii) a reverse PCR primer with an oligonucleotide sequence of SEQ ID NO:2, (iv) a probe with an oligonucleotide sequence of SEQ ID NO:7, 8 and 9, labelled with a fluorophore and a quencher molecule and b) detection of a change of fluorescence the wavelength thereof determining the HPV type 18, 52 and 59; Reaction 2: a) amplification of the nucleic acid fragments of HPV 39, 51, 56 and 66 in the presence of: (i) a nucleic acid polymerase (ii) a forward PCR primer with an oligonucleotide sequence of SEQ ID NO:1, (iii) a reverse PCR primer with an oligonucleotide sequence of SEQ ID NO:3, (iv) a probe with an oligonucleotide sequence of SEQ ID NO: 10, 11, 12 and 13, labelled with a fluorophore and a quencher molecule and b) detection of a change of fluorescence the wavelength thereof determining the HPV type 39,51,56 and 66; Reaction 3: a) amplification of the nucleic acid fragments of HPV 16, 45 and 58 in the presence of: (i) a nucleic acid polymerase (ii) a forward PCR primer with an oligonucleotide sequence of SEQ ID NO:1, (iii) a reverse PCR primer with an oligonucleotide sequence of SEQ ID NO:4, (iv) a probe with an oligonucleotide sequence of SEQ ID NO:14, 15 and 16, labelled with a fluorophore and a quencher molecule and b) detection of a change of fluorescence the wavelength thereof determining the HPV type 16, 45 and 58; Reaction 4: a) amplification of the nucleic acid fragments of HPV 31, 33 and 35 and internal control in the presence of: (i) a nucleic acid polymerase (ii) a forward PCR primer with an oligonucleotide sequence of SEQ ID NO:1, (iii) a reverse PCR primer with an oligonucleotide sequence of SEQ ID NO:5, (iv) a probe with an oligonucleotide sequence of SEQ ID NO:17, 18, 19 and 20, labelled with a fluorophore and a quencher molecule, and (v) internal control, and b) detection of a change of fluorescence the wavelength thereof determining the HPV type 31,33 and 35 and internal control. |
