Nucleic acid probes and methods for detecting plasmodium parasites

Abstract

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.

What is claimed is: 
     
         1 . A composition suitable for detecting  Plasmodium falciparum  in a sample, said composition comprising one or more of: a nucleic acid sequence consisting of PF1 [SEQ ID NO: 9] or the full-length complementary sequence thereof, or a polymerase chain reaction extension product of PF1 [SEQ ID NO: 9] and a reverse primer, and a label; a nucleic acid sequence consisting of PF3 [SEQ ID NO: 11] or the full-length complementary sequence thereof, or a polymerase chain reaction extension product of PF3 [SEQ ID NO: 11] and a reverse primer, and a label; a nucleic acid sequence consisting of PF4 [SEQ ID NO: 12] or the full-length complementary sequence thereof, or a polymerase chain reaction extension product of PF4 [SEQ ID NO: 12] and a reverse primer, and a label; and a nucleic acid consisting of PF5 [SEQ ID NO: 13] or the full-length complementary sequence thereof, or a polymerase chain reaction extension product of PF5 [SEQ ID NO: 13] and a reverse primer and a label. 
     
     
         2 . The composition of  claim 1 , wherein said composition comprises the nucleic acid sequence consisting of PF1 [SEQ ID NO: 9] or the full-length complementary sequence thereof, and a label. 
     
     
         3 . The composition of  claim 1 , wherein said composition comprises the nucleic acid sequence consisting of PF3 [SEQ ID NO: 11] or the full-length complementary sequence thereof, and a label. 
     
     
         4 . The composition of  claim 1 , wherein said composition comprises the nucleic acid sequence consisting of PF4 [SEQ ID NO: 12] or the full-length complementary sequence thereof, and a label. 
     
     
         5 . The composition of  claim 1 , wherein said composition comprises the nucleic acid sequence consisting of PF5 [SEQ ID NO: 13] or the full-length complementary sequence thereof, and a label. 
     
     
         6 . 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 PF1 [SEQ ID NO: 9] or the full-length complementary sequence thereof, and a label; a nucleic acid sequence consisting of PF3 [SEQ ID NO: 11] or the full-length complementary sequence thereof, and a label; a nucleic acid sequence consisting of PF4 [SEQ ID NO: 12] or the full-length complementary sequence thereof, and a label; and a nucleic acid sequence consisting of PF5 [SEQ ID NO: 13] or the full-length complementary sequence thereof, and a label;   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.   
     
     
         7 . The method of  claim 6  wherein the biological sample is selected from the group consisting of whole blood, cerebrospinal fluid (CSF), bone marrow and tissue sample. 
     
     
         8 . The method of  claim 7  wherein the tissue sample is isolated from a spleen. 
     
     
         9 . The method of  claim 6  wherein said composition comprises the nucleic acid sequence consisting of PF1 [SEQ ID NO: 9] or the full-length complementary sequence thereof, and a label. 
     
     
         10 . The method of  claim 6  wherein said composition comprises the nucleic acid sequence consisting of PF3 [SEQ ID NO: 11] or the full-length complementary sequence thereof, and a label. 
     
     
         11 . The method of  claim 6  wherein said composition comprises the nucleic acid sequence consisting of PF4 [SEQ ID NO: 12] or the full-length complementary sequence thereof, and a label. 
     
     
         12 . The method of  claim 6  wherein said composition comprises the nucleic acid sequence consisting of PF5 [SEQ ID NO: 13] or the full-length complementary sequence thereof, and a label. 
     
     
         13 . 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 PF1 [SEQ ID NO: 9] or the full-length complementary sequence thereof; 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.   
     
     
         14 . The method of  claim 6  wherein the biological sample is selected from the group consisting of whole blood, cerebrospinal fluid (CSF), bone marrow and tissue sample. 
     
     
         15 . 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 PF1 [SEQ ID NO: 9] or the full-length complementary sequence thereof, and a label; a nucleic acid sequence consisting of PF3 [SEQ ID NO: 11] or the full-length complementary sequence thereof, and a label; a nucleic acid sequence consisting of PF4 [SEQ ID NO: 12] or the full-length complementary sequence thereof, and a label; and a nucleic acid sequence consisting of PF5 [SEQ ID NO: 13] or the full-length complementary sequence thereof, and a label; under conditions appropriate for specific hybridization; and   c) detecting specific hybridization as an indication of the presence of  Plasmodium falciparum  in the biological sample.   
     
     
         16 . The method of  claim 6  wherein the biological sample is selected from the group consisting of whole blood, cerebrospinal fluid (CSF), bone marrow and tissue sample.