DK2322663T3 - Fremgangsmåder og sekvenser til påvisning og identificering af methicillin-resistente stammer af Staphylococcus aureus MREJ type v - Google Patents
Fremgangsmåder og sekvenser til påvisning og identificering af methicillin-resistente stammer af Staphylococcus aureus MREJ type v Download PDFInfo
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- DK2322663T3 DK2322663T3 DK10181533T DK10181533T DK2322663T3 DK 2322663 T3 DK2322663 T3 DK 2322663T3 DK 10181533 T DK10181533 T DK 10181533T DK 10181533 T DK10181533 T DK 10181533T DK 2322663 T3 DK2322663 T3 DK 2322663T3
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- C12Q1/6888—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for detection or identification of organisms
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Claims (14)
1. Fremgangsmåde til påvisning af forekomsten af MREJ type i, ii, iii og v methicillin-resistente Staphylococcus aureus (MRSA)-stammer omfattende: a) at bringe en prøve, der skal analyseres for forekomsten af MREJ type i, ii, iii og V MRSA-stammerne, hvor hver af MRSA-stammerne omfatter et Staphylococcus-kassettekromosom mec (SCCmec) element indeholdende et mecA-gen indsat i kromosomalt DNA, hvorved der genereres en polymorf højre ydre forbindelse (MREJ) type i, ii, iii eller v-sekvens, der omfatter sekvenser fra både SCCmeo-elementets højre ydre ende og kromosomalt DNA, der grænser op til den højre ydre ende af SCCmec-elementet, i kontakt med en første primer og en anden primer for hver af MREJ typen i, ii, iii og v, hvor hver første primer hybridiserer med SCCmec-elementets højre ydre ende af en MREJ type i, ii, iii eller v-sekvens udvalgt fra gruppen bestående af: SEQ ID NOs: 1,20 til 25, 41 og 199, og komplementer deraf, for MREJ type
1. SEQ ID NOs: 2, 17 til 19, 26, 40, 173 til 183, 185, 186 og 197, og komplementer deraf, for MREJ type ii, SEQ ID NOs: 4 til 16, 104, 184 og 198, og komplementer deraf, for MREJ type iii, og SEQ ID NOs: 47 til 50, og komplementer deraf, for MREJ type v; og hvor hver anden primer hybridiserer med en kromosomal sekvens af S. aureus med henblik på specifikt at generere en amplikon(er), hvis en sådan MREJ type i, ii, iii eller v MRSA-stamme forefindes i prøven; og b) at påvise forekomsten af amplikonen/amplikonerne.
2. Fremgangsmåde ifølge krav 1, hvor den kromosomale sekvens af S. aureus er orfX.
3. Fremgangsmåde ifølge krav 1 eller 2, hvor fremgangsmåden omfatter anvendelsen af mindst en primer og/eller sonde udvalgt blandt de følgende SEQ ID NOs: 65, 80, 154, 155, 64, 71,72, 73, 74, 75, 76, 70, 103, 130, 132, 158, 159, 59, 62, 126, 127, 128, 129, 131, 200, 201, 60, 61, 63, 32, 83, 84, 160, 161, 162,163 og 164, til påvisning af MREJ type v.
4. Fremgangsmåde ifølge krav 3, omfattende anvendelsen af et primerpar bestående af SEQ ID NOs: 64 og 80.
5. Fremgangsmåde ifølge krav 4, yderligere omfattende anvendelsen af mindst en sonde med en sekvens udvalgt fra gruppen bestående af SEQ ID NOs: 32, 83, 84, 160, 161, 162, 163 og 164.
6. Fremgangsmåde ifølge krav 5, hvor primerne og sonderne har de følgende nukleotidsekvenser: SEQ ID NOs: 64, 80, 84, 163, 164 til påvisning af MREJ type V.
7. Fremgangsmåde ifølge et af kravene 1 til 6, hvor flere primere og/eller sonder anvendes sammen inden for det samme fysiske indelukke.
8. Kit til påvisning af MREJ type i, ii, ill og v MRSA-stammer i en prøve omfattende: a) et første sæt af oligonukleotider, som hybridiserer med SCCmec-elemen-tets højre ydre ende af MREJ type i, ii, iii og v-sekvenser udvalgt fra gruppen bestående af SEQ ID NOs: 1, 20 til 25, 41 og 199, og komplementer deraf, for MREJ type i, SEQ ID NOs: 2, 17 til 19, 26, 40, 173 til 183, 185, 186 og 197, og komplementer deraf, for MREJ type ii, SEQ ID NOs: 4 til 16, 104, 184 og 198, og komplementer deraf, for MREJ type iii, og SEQ ID NOs: 47 til 50, og komplementer deraf, for MREJ type v; og b) et andet oligonukleotid, som hybridiserer med en kromosomal sekvens af S. aureus; hvor oligonukleotiderne af a) og b) muliggør den selektive generering af en amplikon/amplikoner, som omfatter sekvenser fra både SCCmec-elementets højre ydre ende og kromosomalt DNA, der grænser op til den højre ydre ende af MREJ type i, ii, iii og v MRSA-stammer.
9. Kit ifølge krav 8, hvor den specifikke sekvens af det S. aureus kromosomale DNA er orfX.
10. Kit ifølge krav 8 eller 9, hvor det andet oligonukleotid i b) omfatter en sekvens udvalgt fra gruppen bestående af SEQ ID NQs: 32, 59, 60 til 64, 70 til 76, 83, 84, 103, 126 til 132, 160 til 164, 200 og 201.
11. Kit ifølge krav 8 eller 9, omfattende et par af oligonukleotider bestående afSEQ ID NOs:64 og 80.
12. Kit ifølge et af kravene 8 til 11, yderligere omfattende mindst en sonde udvalgt fra gruppen bestående af SEQ ID NOs: 32, 83, 84, 160, 161, 162, 163 og 164.
13. Fremgangsmåde ifølge et af kravene 1 til 7 eller kit ifølge et af kravene 8 til 12, hvor den anden primer har en sekvens som anført i SEQ ID NO: 64.
14. Kit ifølge et af kravene 8 til 13 omfattende primere og sonder med sekvenserne som anført i SEQ ID NOs: 64, 80, 84,163 og 164.
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| CA002348042A CA2348042A1 (en) | 2001-06-04 | 2001-06-04 | Sequences for detection and identification of methicillin-resistant staphylococcus aureus |
| EP09174581A EP2236621B1 (en) | 2001-06-04 | 2002-06-04 | Methods and sequences for detection and identification of methicillin-resistant Staphylococcus aureus |
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