Probes, methods and kits for detection and typing of Helicobacter pylori nucleic acids in biological samples
OwnerWilhelmus Quint, Leendert-Jan Van Doorninventors
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OwnerWilhelmus Quint, Leendert-Jan Van Doorninventors
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The present invention relates to a method for the detection and/or typing of Helicobacter pylori (H. pylori) strains present in a sample comprising the steps of: (i) if need be releasing, isolating or concentrating the polynucleic acids in the sample, (ii) amplifying the polynucleic acids of relevant target regions of the vacA gene and possibly other virulence determinant genes (VDG), with suitable primer pairs, said primers being generally applicable on different H. pylori strains, allowing to amplify said relevant target regions of the VDG preferentially in compatible amplification conditions; (iii) hybridizing the polynucleic acids obtained in (i) or (ii) with a set of at least two VDG-derived probes, under appropriate hybridization and wash conditions, and with at least one of said probes hybridizing to a conserved region of a VDG of H. pylori, and with at least one of said probes hybridizing to a variable region of vacA; (iv) detecting the hybrids formed in step (iii), (v) detecting and/or typing H. pylori strains present in a sample from the differential hybridization signals obtained in step (iv), with said typing being the allele-specific detection of a strain according to the VDG alleles present in that particular H. pylori strain, and the said virulence determinant genes being the genetic elements involved in enabling, determining, and marking of the infectivity and/or pathogenicity of said H. pylori strain. The present invention also relates to probes and primers for doing the same as well as Helicobacter pylori detecting/typing kits. The present invention also discloses novel sequences of VDG, which can be used for designing the above-mentioned primers and probes.
A method for the detection of Helicobacter pylori (H. pylori) present in a sample comprising the steps of:
A probe for use in a method of detecting the presence of H. pylori, said probe comprising a sequence selected from the group consisting of SEQ ID NOS:35-39.
A kit for detecting and/or typing H. pylori strains in a sample liable to contain it, comprising the following components:
An isolated vacA polynucleotide sequence selected from the group consisting of:
A method for the detection and/or typing of Helicobacter pylori (H.pylori) strains present in a sample comprising the steps of:
An oligonucleotide primer, wherein said vacA primer is selected from the group consisting of SEQ ID NOS:
A probe for detection or typing of vacA, said probe selected from the group consisting of SEQ ID NOS:2-11, and 28-39.
A vacA-specific oligonucleotide selected from the group consisting of SEQ ID NOS:
Dependencies are inferred from the claim text.