Certains contenus de cette application ne sont pas disponibles pour le moment.
Si cette situation persiste, veuillez nous contacter àObservations et contact
1. (WO1999054508) PROCEDE RAPIDE ET SENSIBLE DE DETECTION D'$i(HISTOPLASMA CAPSULATUM)
Note: Texte fondé sur des processus automatiques de reconnaissance optique de caractères. Seule la version PDF a une valeur juridique

We claim-

1. An oligonucleotide primer pair comprising sequences specific for the 5.8S rRNA gene of Histoplasma capsulatum, or an oligonucleotide comprising a sequence complementary to one or the other of the primer pair specific for the H. capsulatum 5.8S rRNA gene.

2. An oligonucleotide comprising a sequence given by SEQ ID NO:S or a sequence complementary to that given by SEQ ID NO:S.

3. An oligonucleotide comprising a sequence given by SEQ ID NO:4 or a sequence complementary to that given by SEQ ID NO:4.

4. A method of detecting the presence of H. capsulatum in a sample, said method comprising the steps of:
(a) providing a sample suspected of harboring H. capsulatum,
(b) preparing a first amplified nucleic acid by contacting the sample with a first solution comprising an oligonucleotide primer pair specific for a fungal nucleic acid sequence that comprises the coding sequence for the 5.8S rRNA gene, a first DNA polymerase, and a first mixture of deoxynucleotide triphosphates, in a first buffer, and carrying out a polymerase chain reaction on the first solution resulting therefrom;
(c) preparing a second amplified nucleic acid by contacting all or a portion of the first amplified nucleic acid from step (b) with a second solution comprising an oligonucleotide primer pair specific for the H capsulatum 5.8S rRNA gene, a second DNA polymerase, and a second mixture of deoxynucleotide triphosphates, in a second buffer, and carrying out a polymerase chain reaction on the second solution resulting therefrom; and
(d) detecting the presence of DNA derived from H. capsulatum in the second amplified nucleic acid,
whereby the presence of DNA derived from H. capsulatum in the second amplified nucleic acid indicates that H. capsulatum is present in the sample.

5. The method of claim 4 wherein the sample is an environmental sample comprising soil or a clinical sample from a human subject.

6. The method of claim 4 wherein before step (b) the sample is subjected to partial purification comprising separation of any soil or contaminants contained in the sample from the H. capsulatum nucleic acid.

7. The method of claim 4 wherein the first buffer comprises bovine serum albumin.

8. The method of claim 4 wherein the detecting step comprises gel electrophoresis of the second amplified nucleic acid and detecting any H.
capsulatum-sTpeciiic DNA in the resulting gel.

9. The method of claim 4 wherein the detecting step comprises contacting the second amplified nucleic acid with a labeled probe specific to H capsulatum DNA and detecting the label.

10. The method of claim 4 wherein the oligonucleotide primer pair employed in step (b) is specific for the fungal internal transcribed spacer regions flanking the coding sequence for the 5.8S rRNA gene.

11. A method of detecting the presence of Histoplasma capsulatum in a sample, said method comprising the steps of:
(a) providing a sample suspected of harboring H. capsulatum;
(b) preparing a first amplified nucleic acid by contacting the sample with a first solution comprising an oligonucleotide comprising the sequence given by SEQ ID NO: l, an oligonucleotide comprising the sequence given by SEQ ID NO:2, a first DNA polymerase, and a first mixture of deoxynucleoside
triphosphates, in a first buffer, and carrying out a polymerase chain reaction on the first solution resulting therefrom;
(c) preparing a second amplified nucleic acid by contacting all or a portion of the first amplified nucleic acid from step (b) with a second solution comprising an oligonucleotide comprising the sequence given by SEQ ID NO:S, an oligonucleotide comprising the sequence given by SEQ ID NO:4, a second DNA polymerase, and a second mixture of deoxynucleoside triphosphates, in a second buffer, and carrying out a polymerase chain reaction on the second solution resulting therefrom; and
(d) detecting the presence of DNA derived from H. capsulatum in the second amplified nucleic acid,
whereby the presence of DNA derived from H. capsulatum in the second amplified nucleic acid indicates that H. capsulatum is present in the sample.

12. The method of claim 1 1 wherein the sample is an environmental sample comprising soil or a clinical sample from a human subject.

13. The method of claim 11 wherein before step (b) the sample is subjected to partial purification comprising separation of any soil or contaminants contained in the sample from the H. capsulatum nucleic acid.

14. The method of claim 11 wherein the first buffer comprises bovine serum albumin.

15. The method of claim 11 wherein the detecting step comprises gel electrophoresis of the second amplified nucleic acid and detecting any H.
capsulatum-speci c DNA in the resulting gel.

16. The method of claim 11 wherein the detecting step comprises contacting the second amplified nucleic acid with a labeled probe specific to H. capsulatum DNA and detecting the label.