Claims
- 1. A synthetic oligonucleotide useful as an amplifier probe in a sandwich hybridization assay for Chlamydia trachomatis consisting of:
- (a) a first segment having a nucleotide sequence complementary to a segment of the Chlamydia pCHL2 plasmid,
- (b) a second segment having a nucleotide sequence complementary to an oligonucleotide unit of a nucleic acid multimer but not complementary to Chlamydia trachomatis nucleic acid, and
- (c) optionally one or more non-complementary segments each consisting of a nucleotide sequence that is not complementary to Chlamydia trachomatis nucleic acid,
- wherein the first segment has a sequence selected from the group consisting of ##STR14##
- 2. A synthetic oligonucleotide useful as a capture probe in a sandwich hybridization assay for Chlamydia trachomatis consisting of:
- (a) a first segment having a nucleotide sequence complementary to a segment of the Chlamydia pCHL2 plasmid,
- (b) a second segment having a nucleotide sequence complementary to an oligonucleotide bound to a solid phase but not complementary to Chlamydia trachomatis nucleic acid, and
- (c) optionally one or more non-complementary segments each consisting of a nucleotide sequence that is not complementary to Chlamydia trachomatis nucleic acid, wherein the first segment has a sequence selected from the group consisting of ##STR15##
- 3. A solution-phase sandwich hybridization assay for detecting Chlamydia trachomatis DNA in an analyte, the assay comprising the steps of:
- (a) contacting the analyte under specific hybridizing conditions with an excess of: (i) an amplifier probe oligonucleotide consisting of a first segment having a nucleotide sequence complementary to a segment of the Chlamydia pCHL2 plasmid, a second segment having a nucleotide sequence complementary to an oligonucleotide unit of a nucleic acid multimer but not complementary to Chlamydia trachomatis nucleic acid, and optionally one or more non-complementary segments each consisting of a nucleotide sequence that is not complementary to Chlamydia trachomatis nucleic acid; and (ii) a capture probe oligonucleotide consisting of a first segment having a nucleotide sequence complementary to another segment of the Chlamydia pCHL2 plasmid, a second segment having a nucleotide sequence complementary to an oligonucleotide bound to a solid phase but not complementary to Chlamydia trachomatis nucleic acid, and optionally one or more non-complementary segments each consisting of a nucleotide sequence that is not complementary to Chlamydia trachomatis nucleic acid;
- (b) contacting under hybridizing conditions the product of step (a) with the oligonucleotide bound to the solid phase;
- (c) thereafter separating materials not bound to the solid phase;
- (d) contacting under hybridizing conditions the solid phase complex product of step (c) with the nucleic acid multimer, said multimer comprising (i) at least one oligonucleotide unit that is complementary to the second segment of the amplifier probe oligonucleotide and (ii) a multiplicity of second oligonucleotide units that are complementary to a labeled oligonucleotide;
- (e) removing unbound multimer;
- (f) contacting under hybridizing conditions the solid phase complex product of step (e) with the labeled oligonucleotide;
- (g) removing unbound labeled oligonucleotide; and
- (h) detecting the presence of Chlamydia trachomatis by detecting the presence of label in the solid phase complex product of step (g),
- wherein the first segment of the amplifier probe oligonucleotide has a sequence selected from the group consisting of ##STR16## and the first segment of the capture probe oligonucleotide has a sequence selected from the group consisting of ##STR17##
CROSS-REFERENCE TO RELATED APPLICATIONS
This application is a continuation of application Ser. No. 07/824,795, filed Jan. 22, 1992, now abandoned, which application is a division of U.S. patent application Ser. No. 340,031, filed Apr. 18, 1989 now U.S. Pat. No. 5,124,246, which is in turn a continuation-in-part of U.S. patent application Ser. No. 252,638, filed 30 Sep. 1988, now abandoned, which is in turn a continuation-in-part of U.S. patent application Ser. No. 185,201, filed 22 Apr. 1988, now abandoned, which is in turn a continuation-in-part of U.S. patent application Ser. No. 109,282, filed 15 Oct. 1987, now abandoned.
US Referenced Citations (8)
Foreign Referenced Citations (8)
Number |
Date |
Country |
0154884 |
Sep 1985 |
EPX |
0225807 |
Jun 1987 |
EPX |
0317077 |
May 1989 |
EPX |
2125964 |
Mar 1984 |
GBX |
WO8706270 |
Oct 1987 |
WOX |
WO8904375 |
May 1989 |
WOX |
WO8903891 |
May 1989 |
WOX |
WO9002819 |
Mar 1990 |
WOX |
Non-Patent Literature Citations (6)
Entry |
Palmer et al. (1986) Plasmid, vol. 16, pp. 52-62. |
Chu et al., Nucleic Acids Research (1986) 14(14):5591-5603. |
Synanen et al., Nucleic Acids Research (1986) 14(12):5037-5048. |
Syvanen et al., Nucleic Acids Research (1988) 16(23):11327-11338. |
Fahrlander et al., "Amplifying DNA probe signals: a `Christmas tree` approach" Bio/Technology (1988) 6:1165-1168. |
Ziemer et al., "Sequence of Hepatitis B virus DNA incorporated into the genome of human hepatoma cell line" Virology (1985) 53:(3):885-892. |
Divisions (1)
|
Number |
Date |
Country |
Parent |
340031 |
Apr 1989 |
|
Continuations (1)
|
Number |
Date |
Country |
Parent |
824795 |
Jan 1992 |
|
Continuation in Parts (3)
|
Number |
Date |
Country |
Parent |
252638 |
Sep 1988 |
|
Parent |
185201 |
Apr 1988 |
|
Parent |
109282 |
Oct 1987 |
|