EP0327390A2
Expired / Lapsed
EP
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Probe for pneumocystis carinii
Assignee: Oxford University Innovation Ltd
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Filing: 1989-02-03
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Granted: —
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Expires: 2009-02-03
Abstract
Abstract The invention provides DNA probes which hybridize to the DNA of Pneumocystis carinii but not to mammalian DNA, and which form the basis of an in vitro diagnostic test for viable P.carinii organisms in respiratory secretion or even sputum. The probes are made by recombinant DNA technology from infected rat lung tissue. Preferred in the plasmid designated pAZ112, derived from the vector pUC13, having a size of approximately 9.8 kbp and which has been partly sequenced.
Inventors
- 👤 Ann Elizabeth Wakefield
- 👤 Julian Meurglyn Hopkin
- 👤 Edward Richard Moxon
Bibliographic Details
Application Number
—
Jurisdiction
EP
Data Source
google_patents
Last Synchronized
2026-11-03 00:08
CPC Classifications
C; C07; C07K; C07K14/00; C07K14/37; C12; C12Q; C12Q1/00; C12Q1/68; C12Q1/6876; C12Q1/6888; C12Q1/6895
Claims Analysis
Total Claims: 19
Claim 1
Independent Claim
21 words
A DNA probe produced by recombinant DNA technology which hybridizes to the DNA of Pneumocystis carinii but not to mammalian DNA.
Claim 2
Dependent Claim
21 words
A DNA probe produced by recombinant DNA technology which hybridizes to the DNA of Pneumocystis carinii but not to mammalian DNA.
Claim 3
Dependent Claim
20 words
A probe as claimed in claim 1 which comprises part or all of the insert of the plasmid designated pAZ101.
Claim 4
Dependent Claim
20 words
A probe as claimed in claim 1 which comprises part or all of the insert of the plasmid designated pAZ102.
Claim 5
Dependent Claim
20 words
A probe as claimed in claim 1 which comprises part or all of the insert of the plasmid designated pAZ112.
Claim 6
Dependent Claim
56 words
A probe as claimed in claim 4, wherein the insert has a length of approximately 7.1 kbp and includes the following sites for the following restriction endonucleases: Restriction endonuclease Minimum number of sites in insert Hae III 2 Dra I 9 Hind III 1 EcoR I 3 Xba I 3 ApaI, SmaI, PstI, PvuII, SalI 0
Claim 7
Dependent Claim
26 words
A probe as claimed in claim 4 or claim 5, wherein the insert includes a DNA sequence which is substantially the sequence shown in Figure 2.
Claim 8
Dependent Claim
43 words
A DNA probe which hybridizes to the DNA of P.carinii but not to mammalian DNA, which probe includes a sequence of at least 15 nucleotides which has at least 70% homology with the sequence shown in Figure 2 or with the complementary sequence.
Claim 9
Dependent Claim
43 words
A DNA probe which hybridizes to the DNA of P.carinii but not to mammalian DNA, which probe includes a sequence of at least 15 nucleotides which has at least 70% homology with the sequence shown in Figure 2 or with the complementary sequence.
Claim 10
Dependent Claim
32 words
A probe as claimed in any one of claims 1 to 7, which hybridizes to a DNA sequence that is present in more than one copy in the genome of P.carinii .
Claim 11
Dependent Claim
22 words
A probe as claimed in any one of claims 1 to 8, wheron at least one marker atom or group is present.
Claim 12
Dependent Claim
17 words
A micro-organism which is capable of producing the DNA of any one of claims 1 to 8.
Claim 13
Independent Claim
119 words
A method of making a DNA probe for Pneumocystis carinii , which method comprises the steps of - extracting P.carinii from infected lung tissue of a mammal, - recovering DNA from the P.carinii , inserting the DNA by ligation into a vector and using the resulting recombinant vectors to transform microorganisms, - screening the transformed micro-organisms for hybridization to DNA of the mammal, selecting ones which do not hybridise, and extracting recombinant vectors from the selected micro-organisms, - screening the extracted recombinant vectors by in situ hybridization with lung tissue of a mammal infected with P.carinii , and selecting recombinant vectors that do hybridize, to the P.carinii , and - culturing the micro-organisms that produce the selected recombinant vectors.
Claim 14
Independent Claim
119 words
A method of making a DNA probe for Pneumocystis carinii , which method comprises the steps of - extracting P.carinii from infected lung tissue of a mammal, - recovering DNA from the P.carinii , inserting the DNA by ligation into a vector and using the resulting recombinant vectors to transform microorganisms, - screening the transformed micro-organisms for hybridization to DNA of the mammal, selecting ones which do not hybridise, and extracting recombinant vectors from the selected micro-organisms, - screening the extracted recombinant vectors by in situ hybridization with lung tissue of a mammal infected with P.carinii , and selecting recombinant vectors that do hybridize, to the P.carinii , and - culturing the micro-organisms that produce the selected recombinant vectors.
Claim 15
Dependent Claim
24 words
A method as claimed in claim 11, wherein the mammal is a rat, and the P.carinii is separated from rat lung tissue by filtration.
Claim 16
Dependent Claim
25 words
A method as claimed in claim 11 or claim 12, wherein the vector is a plasmid and the micro-organisms are of the species E. coli.
Claim 17
Independent Claim
24 words
A method as claimed in any one of claims 11 to 13, wherein the in situ hybridization step is performed using an enzyme label.
Claim 18
Dependent Claim
54 words
An in vitro method of diagnosing Pneumocystis carinii infection, which method comprises contacting DNA, from respiratory secretion of a patient suspected of being infected, under hybridizing conditions with the probe claimed in any one of claims 1 to 9 or the probe made by the method of any one of claims 11 to 14.
Claim 19
Dependent Claim
15 words
An in vitro method as claimed in claim 15, wherein the respiratory secretion is sputum.
References Cited (Backward Citations)
No citation records indexed.
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