Primers and probes for detecting mycobacteria
OwnerClothilde Fernandez, Marine Mariusinventors
Life of this patent
Estimating the term…
Abstract
The present invention is directed to a set of primers for amplifying by Polymerase Chain Reaction (PCR) a sequence of any mycobacterial rrs gene present in a sample, wherein said set comprises at least three pairs of primers and allows the amplification of a sequence from the rrs gene of any mycobacterial species. The invention is also directed to probes, associated to the pairs of primers, allowing the specific detection of the mycobacterial rrs sequences amplified by the pairs of primers. The invention also concerns different methods and uses of these primers and probes, in order to exhaustively and specifically detect mycobacterial presence in any product, especially pharmaceutical products such as vaccines.
Claims20
A set of primers for amplifying by Polymerase Chain Reaction (PCR) a sequence of any mycobacterial rrs gene present in a sample, wherein said set comprises at least the three following pairs of primers:
- 1st Pair:
- Forward primer F1 having the sequence:
- (SEQ ID NO:
- 1)
- 5′-CCTGGTAGTCCACGCCGTAA-3′
- or
- modifications thereof,
- Reverse primer R1 having the sequence:
- (SEQ ID NO:
- 2)
- 5′-CGGATCCCAAGGAAGGAAAC-3′
- or
- modifications thereof;
- 2nd Pair:
- Forward primer F3 having the sequence:
- (SEQ ID NO:
- 4)
- 5′-CACAGGACGCCGGTAGAGAT-3′
- or
- modifications thereof,
- Reverse primer R3 having the sequence:
- (SEQ ID NO:
- 5)
- 5′-CATGCACCACCTGCACACA-3′
- or
- modifications thereof;
- 3rd Pair:
- Forward primer F4 having the sequence:
- (SEQ ID NO:
- 7)
- 5′-CCCTTATGTCCAGGGCTTCA-3′
- or
- modifications thereof,
- Reverse primer R4 having the sequence:
- (SEQ ID NO:
- 8)
- 5′-GGCATCGCAGCCCTTTG-3′
- or
- modifications thereof;
- wherein said modifications are the addition and/or the deletion of 1 to 4 nucleotides, at the 3′ and/or 5′ terminus, and/or the substitution of 1 or 2 nucleotides, and
- wherein said three pairs of primers allow the amplification of a sequence from the rrs gene of any mycobacterial species.
A method for amplifying and detecting sequences of the rrs gene of any mycobacteria present in a sample, comprising the following steps, carried out simultaneously or sequentially:
- a) Carrying out in vitro PCR on the nucleic acids extracted from said sample with a set of primers;
- and
- b) Detecting the presence or absence of an amplified product,
- wherein the set of primers comprises at least the three following pairs of primers:
- 1st Pair:
- Forward primer F1 having the sequence:
- 5′-CCTGGTAGTCCACGCCGTAA-3′
- (SEQ ID NO:
- 1) or modifications thereof.
- Reverse primer R1 having the sequence:
- 5′-CGGATCCCAAGGAAGGAAAC-3′
- (SEQ ID NO:
- 2) or modifications thereof;
- 2nd Pair:
- Forward primer F3 having the sequence:
- 5′-CACAGGACGCCGGTAGAGAT-3′
- (SEQ ID NO:
- 4) or modifications thereof,
- Reverse primer R3 having the sequence:
- 5′-CATGCACCACCTGCACACA-3′
- (SEQ ID NO:
- 5) or modifications thereof;
- 3rd Pair:
- Forward primer F4 having the sequence:
- 5′-CCCTTATGTCCAGGGCTTCA-3′
- (SEQ ID NO:
- 7) or modifications thereof,
- Reverse primer R4 having the sequence:
- 5′-GGCATCGCAGCCCTTTG-3′
- (SEQ ID NO:
- 8) or modifications thereof;
- wherein said modifications are the addition and/or the deletion of 1 to 4 nucleotides, at the 3′ and/or 5′ terminus, and/or the substitution of 1 or 2 nucleotides, and
- wherein said three pairs of primers allow the amplification of a sequence from the rrs gene of any mycobacterial species.
Dependencies are inferred from the claim text.