Nucleic acid probes and methods for detecting plasmodium parasites
Inventors
Shah, Jyotsna S. • Wronska, Danuta • Weltman, Helena • Harris, Nick S. • SCHERINI-WARD, Suzanne • MARK, Oliva
Assignees
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Publication Number
US-8592568-B2
Publication Date
2013-11-26
Expiration Date
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.
Core Innovation
The disclosed invention concerns nucleic-acid probe sets for hybridization-based detection and selective discrimination of Plasmodium species in a sample. The probes comprise labeled DNA/RNA/PNA nucleic-acid probes used for hybridization analysis to distinguish Plasmodium falciparum, Plasmodium vivax, Plasmodium malariae, and Plasmodium ovale based on hybridization patterns.
The probe sets define genus probes, including PGenus1 and PGenus2, and species-specific probes, including PV1, PV2, PF1-PF5, PM1, and PO1. The probes are described as including full-length complementary sequences and shorter contiguous subsequences, with contiguous nucleotides described as ≤5/10/13/15 nt, for detection methods.
Experimental dot-blot analysis is reported to show expected positive and negative hybridization patterns across the Plasmodium species, consistent with selective discrimination. The document further lists specific probe sequences for many probes, including PV1, PV2, and PF2, and indicates the use of labeled probes in the hybridization assay to obtain the expected results.
Claims Coverage
The independent claim covers a Plasmodium detection composition defined by a labeled probe set. The coverage is centered on three labeled probe options, PV1, PV2, and PF2, each used as a probe or its full-length complementary sequence, with dependent claims narrowing to specific probe selections and target species.
Labeled PV1 probe or full-length complementary sequence for Plasmodium detection
A composition suitable for detecting Plasmodium in a sample, comprising a probe consisting of PV1 [SEQ ID NO.: 7] or the full-length complementary sequence thereof, and a label.
Labeled PV2 probe or full-length complementary sequence for Plasmodium detection
A composition suitable for detecting Plasmodium in a sample, comprising a probe consisting of PV2 [SEQ ID NO.: 8] or the full-length complementary sequence thereof, and a label.
Labeled PF2 probe or full-length complementary sequence for Plasmodium detection
A composition suitable for detecting Plasmodium in a sample, comprising a probe consisting of PF2 [SEQ ID NO.: 10] or the full-length complementary sequence thereof, and a label.
Plasmodium vivax detection using PV1 and PV2 labeled probes
A composition for detecting Plasmodium vivax that includes probes consisting of PV1 [SEQ ID NO.: 7] and PV2 [SEQ ID NO.: 8] or their full-length complementary sequences, each with a label.
Plasmodium falciparum detection using PF2 labeled probe
A composition for detecting Plasmodium falciparum that includes a probe consisting of PF2 [SEQ ID NO.: 10] or its full-length complementary sequence together with a label.
Overall, the claim set defines a Plasmodium detection composition based on labeled nucleic-acid probes PV1, PV2, and PF2 or their full-length complementary sequences, with narrower dependent coverage for detecting Plasmodium vivax using PV1 and PV2, and detecting Plasmodium falciparum using PF2.
Stated Advantages
Selective discrimination of Plasmodium species based on hybridization-based detection patterns.
Expected positive/negative hybridization patterns are demonstrated by dot-blot analysis for the probe sets.
Documented Applications
Hybridization assay/dot-blot analysis for detecting Plasmodium species (P. falciparum, P. vivax, P. malariae, P. ovale) in a sample using labeled nucleic-acid probes.
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