20080241891 | Method for the Amplification and Detection of HBV DNA Using a Transcription Based Amplification - The present invention provides a method for the transcription based amplification of a target HBV nucleic acid sequence starting from HBV DNA optionally present in a sample, comprising the steps of, —incubating the sample, suspected to contain HBV, in an amplification buffer with one or more restriction enzymes capable of cleaving the HBV DNA at a selected restriction site, said restriction enzyme creating a defined 3′ end of said HBV DNA stand(s), a promoter-primer, said promoter-primer having a 5′ region comprising the sequence of a promoter recognized by a DNA-dependent RNA polymerase and a 3′ region complementary to the define 3′ end of the DNA strand, a second or reverse primer, having the opposite polarity of the promoter-primer and comprising the 5′ end of the said target sequence, and in case of HBV ssDNA as the target sequence, a restriction primer, maintaining the thus created reaction mixture under the appropriate conditions for a sufficient amount of time for a digestion by the restriction enzyme to take place, subjecting the sample thus obtained to a heat treatment at a temperature and time sufficient to inactivate the restriction enzyme and/or to render at least partially a double strand single stranded, adding the following reagents to the sample: an enzyme having RNA dependent DNA polymerase activity, and maintaining the thus created reaction mixture under the appropriate conditions to a sufficient amount of time for the amplification to take place. | 10-02-2008 |
20080280284 | Nucleic Acid Sequences That Can Be Used as Primers and Probes in the Amplification and Detection of Hsv Dna and Method for the Amplification and Detection of Hsv Dna Using a Transcription Based Amplification - The present invention is related to a pair of oligonucleotide primers for the amplification of HSV nucleic acid comprising: a) an oligonucleotide, 10-50 nucleotides in length, preferably 10-35 nucleotides in length, comprising at least a fragment of 10 nucleotides of a sequence selected from the group consisting of: 5′-ACGTTCACCAAGCTGCTGCT-3′, or its complementary sequence and b) an oligonucleotide, 10-50 nucleotides in length, preferably 10-35 nucleotides in length, comprising at least a fragment of 10 nucleotides of a sequence selected from the group consisting of: 5′CCAGGGCCCTGGAGGTGCGG-3′, or its complementary sequence. The invention also relates to probes, method for amplifying an HSV DNA target, method of specific ou aspecific detection of HSV type 1 and 2 and test kit to do possible the detection of HSV. The present invention is especially useful in methods for practicing nucleic acid test. | 11-13-2008 |