Primers and probes for detection and discrimination of Ebola virus
This invention relates to primers and probes for detecting Ebola virus and one or more subtypes of Ebola virus as well as kits including the probes and primers and methods of using the probes and primers.
1. A method of determining the presence of a viral hemorrhagic fever virus nucleic acid in a sample comprising:
a) contacting the sample with at least one probe selected from a group consisting of SEQ ID:21, SEQ ID NO:22, SEQ ID NO: 23, SEQ ID NO:24, SEQ ID NO:25, SEQ ID NO:26, SEQ ID NO: 27, SEQ ID NO:28, SEQ ID NO:29, or SEQ ID:30 capable of hybridizing under stringent conditions to the viral hemorrhagic fever nucleic acid; and detecting hybridization between the viral hemorrhagic fever nucleic acid and the probe, wherein the detection of hybridization indicates the presence of the viral hemorrhagic fever virus nucleic acid in the sample;
wherein the viral hemorrhagic fever virus nucleic acid is an Ebola virus nucleic acid; and wherein the Ebola virus nucleic acid is SEQ ID NO: 37.
2. The method of claim 1 , wherein the probe is labeled.
3. The method of claim 2 , wherein the probe is radiolabeled, or fluorescently labeled.
4. The method of claim 2 , wherein the probe is labeled with FAM and fluorescent quencher.
5. The method of claim 1 , further comprising amplifying the viral hemorrhagic fever nucleic acid by polymerase chain reaction (PCR), reverse transcriptase polymerase PCR (RT-PCR), or real time reverse transcriptase PCR (rt RT-PCR).
6. The method of claim 5 , wherein the amplifying employs primers that consist of the nucleotide sequences set forth as SEQ ID NO: 1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10, SEQ ID NO:11, SEQ ID NO:12, SEQ ID NO: 13, SEQ ID NO:14, SEQ ID NO:15, SEQ ID NO:16, SEQ ID NO:17, SEQ ID NO: 18, SEQ ID NO: 19, or SEQ ID NO:20.