The United States of America As Represented by the Department of Health

The United States of America, As Represented by the Secretary, Department of Health and Human Services

University of Cincinnati

The United States of America

The Government of the United States of America, As Represented by the

The United States of America As Represented by the Secretary of Health

Centers for Disease Control and Prevention

Timothy J Lott

Errol Reiss

Christine J Morrison

Brent Lasker

Sandra Zakroff

Leonard W Mayer

Rosely M Zancope-Oliveira

George S Deepe

Cheryl M Elie

The present invention provides the nucleotide sequence of the M antigen gene of, which is set forth in the Sequence Listing as SEQ ID NO:1, a nucleic acid having the nucleotide sequence complementary thereto, and nucleic acids having a nucleotide sequence which is substantially the same as the foregoing nucleotide sequences. The present invention also provides vectors and host expressions systems containing the foregoing nucleic acids, and isolated or recombinantly-produced antigens encoded by the foregoing nucleic acids. The present invention further provides antibodies generated against the foregoing antigens, and methods and kits for detecting a previous or currentinfection in a subject, and for diagnosing histoplasmosis.

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The nucleic acid sequence encoding the internal transcribed spacer 2 region of Candida, the organism causing candidiasis, for various Candida species. Nucleic acid molecules useful as probes for detecting Candida species are described. The nucleic acid molecules are useful in methods for the detection and diagnosis of Candida infection in a sample or subject.

Nucleic acid probes for detecting Candida species

Provided is an isolated double-stranded nucleic acid consisting essentially of the nucleotide sequences defined in the Sequence Listing by SEQ ID Nos:5-9. These are the ITS2 sequences for C. albicans, C. parapsilosis, C. tropicalis, C. glabrata and C. krusei. A method of diagnosing systemic candidiasis in a subject is also provided. The method comprises the steps of: (a) collecting blood from the subject into tubes containing detergent, polypropylene glycol, sodium poyantholesulfonate, and sodium ethylene diamine tetraacetic acid; (b) lysing Candida cells using ZYMOLYASE.RTM.-100T.TM. enzyme mixture with agitation; (c) extracting and precipitating the DNA from the lysed cells; (d) amplifying the precipitated DNA using universal fungal primer pairs derived from the internal transcribed spacer regions of the Candida ribosomal DNA; and (e) detecting amplified DNA from Candida by hybridizing the amplified DNA with a probe that selectively hybridizes with Candida DNA, the presence of amplified DNA indicating systemic candidiasis.

Nucleic acid probes for candida parapsilosis and methods for detecting

Provided is an isolated double-stranded nucleic acid consisting essentially of the nucleotide sequences defined in the Sequence Listing by SEQ ID Nos: 5-9. These are the ITS2 sequences for C. albicans, C. parapsilosis, C. tropicalis, C. glabrata and C. krusei. A method of diagnosing systemic candidiasis in a subject is also provided. The method comprises the steps of: (a) collecting blood from the subject into tubes containing detergent, polypropylene glycol, sodium poyantholesulfonate, and sodium ethylene diamine tetraacetic acid; (b) lysing Candida cells using ZYMOLYASE.RTM.-100T with agitation; (c) extracting and precipitating the DNA from the lysed cells; (d) amplifying the precipitated DNA using universal fungal primer pairs derived from the internal transcribed spacer regions of the Candida ribosomal DNA; and (e) detecting amplified DNA from Candida by hybridizing the amplified DNA with a probe that selectively hybridizes with Candida DNA, the presence of amplified DNA indicating systemic candidiasis.

Nucleic acid sequences and methods for detecting candida tropicalis in

Provided is an isolated double-stranded nucleic acid consisting essentially of the nucleotide sequences defined in the Sequence Listing by SEQ ID Nos:5-9. These are the ITS2 sequences for C. albicans, C. parapsilosis, C. tropicalis, C. glabrata and C. krusei. A method of diagnosing systemic candidiasis in a subject is also provided. The method comprises the steps of: (a) collecting blood from the subject into tubes containing detergent, polypropylene glycol, sodium poyantholesulfonate, and sodium ethylene diamine tetraacetic acid; (b) lysing Candida cells using ZYMOLYASE.RTM.-100T with agitation; (c) extracting and precipitating the DNA from the lysed cells; (d) amplifying the precipitated DNA using universal fungal primer pairs derived from the internal transcribed spacer regions of the Candida ribosomal DNA; and (e) detecting amplified DNA from Candida by hybridizing the amplified DNA with a probe that selectively hybridizes with Candida DNA, the presence of amplified DNA indicating systemic candidiasis.

Nucleic acid probes and methods for detecting Candida krusei cells in

Provided is an isolated double-stranded nucleic acid consisting essentially of the nucleotide sequences defined in the Sequence Listing by SEQ ID NOs: 5-9. These are the ITS2 sequences for C. albicans, C. parapsilosis, C. tropicalis, C. glabrata and C. krusei. A method of diagnosing systemic candidiasis in a subject is also provided. The method comprises the steps of: (a) collecting blood from the subject into tubes containing detergent, polypropylene glycol, sodium poyantholesulfonate, and sodium ethylene diamine tetraacetic acid; (b) lysing Candida cells using ZYMOLYASE.RTM.-100T with agitation; (c) extracting and precipitating the DNA from the lysed cells; (d) amplifying the precipitated DNA using universal fungal primer pairs derived from the internal transcribed spacer regions of the Candida ribosomal DNA; and (e) detecting amplified DNA from Candida by hybridizing the amplified DNA with a probe that selectively hybridizes with Candida DNA, the presence of amplified DNA indicating systemic candidiasis.

Nucleic acid probes and methods for detecting Candida glabrata DNA in

Provided is an isolated double-stranded nucleic acid consisting essentially of the nucleotide sequence defined in the Sequence Listing by SEQ ID NO:5. This is the C. albicans ITS2 sequence and includes a nucleic acid comprising a nucleotide sequence that is specific for Candida albicans. Further examples of an isolated double stranded nucleic acid of the present invention consist essentially of the nucleotide sequences defined in the Sequence Listing by SEQ ID NOs:6-9. These are the ITS2 sequences for C. parapsilosis, C. tropicalis, C. glabrata and C. krusei. A method of diagnosing systemic candidiasis in a subject is also provided. The method comprises the steps of: (a) collecting blood from the subject into tubes containing detergent, polypropylene glycol, sodium polyanetholesulfonate, and sodium ethylene diamine.RTM. tetraacetic acid; (b) lysing Candida cells using ZYMOLYASE.RTM.-100T with agitation; (c) extracting and precipitating the DNA from the lysed cells; (d) amplifying the precipitated DNA using universal fungal primer pairs derived from the internal transcribed spacer regions of the Candida ribosomal DNA; and (e) detecting amplified DNA from Candida by hybridizing the amplified DNA with a probe that selectively hybridizes with Candida DNA, the presence of amplified DNA indicating systemic candidiasis.

Growing community of inventors

Atlanta, GA, United States of America

Timothy J Lott