Nucleic acid probes and methods for detecting plasmodium parasites
This invention relates to novel nucleic acid probes and methods for detecting Plasmodium parasites as well as detecting different Plasmodium parasites selectively from one another.
1. A composition suitable for detecting Plasmodium falciparum in a sample, said composition comprising one or more of: a labeled nucleic acid sequence consisting of PF4 [SEQ ID NO: 12] or the labeled full-length complementary sequence thereof; and a labeled nucleic acid sequence consisting of PF5 [SEQ ID NO: 13] or the labeled full-length complementary sequence thereof; wherein any said label is selected from one or more of radioisotopes, biotin, digoxigenin, fluorochromes, modified nucleotides and enzymes.
2. The composition of claim 1 , wherein said composition comprises the labeled nucleic acid sequence consisting of PF4 [SEQ ID NO: 12] or the labeled full-length complementary sequence thereof.
3. The composition of claim 1 , wherein said composition comprises the labeled nucleic acid sequence consisting of PF5 [SEQ ID NO: 13] or the labeled full-length complementary sequence thereof.
4. A method for detecting Plasmodium falciparum in a biological sample, the method comprising:
a) providing a biological sample to be analyzed for the presence of Plasmodium falciparum;
b) extracting nucleic acid from the biological sample;
c) providing a composition suitable for detecting Plasmodium falciparum in a sample, said composition comprising one or more of: a nucleic acid sequence consisting of a labeled PF3 [SEQ ID NO: 11] or the labeled full-length complementary sequence thereof; a nucleic acid sequence consisting of a labeled PF4 [SEQ ID NO: 12] or the labeled full-length complementary sequence thereof; and a nucleic acid sequence consisting of a labeled PF5 [SEQ ID NO: 13] or the labeled full-length complementary sequence thereof;
d) contacting the extracted nucleic acid from step b) with the composition suitable for detecting Plasmodium falciparum of step c) under conditions appropriate for detecting specific hybridization;
e) detecting specific hybridization in step d) as an indication of the presence of Plasmodium falciparum in the biological sample.
5. The method of claim 4 wherein the biological sample is selected from the group consisting of whole blood, cerebrospinal fluid (CSF), bone marrow and tissue sample.
6. The method of claim 5 wherein the tissue sample is isolated from a spleen.
7. The method of claim 4 wherein said composition comprises the nucleic acid sequence consisting of the labeled PF3 [SEQ ID NO: 11] or the labeled full-length complementary sequence thereof.
8. The method of claim 4 wherein said composition comprises the nucleic acid sequence consisting of the labeled PF4 [SEQ ID NO: 12] or the labeled full-length complementary sequence thereof.
9. The method of claim 4 wherein said composition comprises the nucleic acid sequence consisting of the labeled PF5 [SEQ ID NO: 13] or the labeled full-length complementary sequence thereof.
10. A method for detecting Plasmodium falciparum in a biological sample, the method comprising:
a) providing a biological sample to be analyzed for the presence of Plasmodium falciparum;
b) extracting nucleic acid from the biological sample;
c) providing a composition suitable for detecting Plasmodium falciparum in a sample, said composition comprising one or more of: a nucleic acid sequence consisting of PF3 [SEQ ID NO: 11] or the full-length complementary sequence thereof; a nucleic acid sequence consisting of PF4 [SEQ ID NO: 12] or the full-length complementary sequence thereof; and a nucleic acid sequence consisting of PF5 [SEQ ID NO: 13] or the full-length complementary sequence thereof;
d) contacting the extracted nucleic acid from step b) with the composition suitable for detecting Plasmodium falciparum of step c) under conditions appropriate for specific hybridization and extension of hybridized nucleic acids by polymerase chain reaction in the presence of a label;
e) detecting polymerase chain reaction extension products containing label as an indication of the presence of Plasmodium falciparum in the biological sample.
11. The method of claim 10 wherein the biological sample is selected from the group consisting of whole blood, cerebrospinal fluid (CSF), bone marrow and tissue sample.
12. A method for detecting the presence of Plasmodium falciparum in a biological sample, the method comprising the steps of:
a) providing a biological sample;
b) contacting the biological sample with a composition suitable for detecting Plasmodium falciparum in a hybridization assay, said composition comprising one or more of: a nucleic acid sequence consisting of a labeled PF3 [SEQ ID NO: 11] or the labeled full-length complementary sequence thereof; a nucleic acid sequence consisting of a labeled PF4 [SEQ ID NO: 12] or the labeled full-length complementary sequence thereof; and a nucleic acid sequence consisting of a labeled PF5 [SEQ ID NO: 13] or the labeled full-length complementary sequence thereof; under conditions appropriate for specific hybridization; and
c) detecting specific hybridization as an indication of the presence of Plasmodium falciparum in the biological sample.
13. The method of claim 12 wherein the biological sample is selected from the group consisting of whole blood, cerebrospinal fluid (CSF), bone marrow and tissue sample.