Oligonucleotides and process for detection of swine flu virus
AssigneeDefence Research & Development Organisation
InventorsManmohan Parida, Jyoti Shukla, Santhosh Sannarangiah, Sashi Sharma, Venkata Lakshmana Putcha Rao, Rajagopalan Vijayaraghavan
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Abstract
A process of detection of pandemic swine flu virus H1N1 type in a sample is provided herein. Also provided are highly specific oligonucleotides useful for rapid detection of swine flu virus in a sample, as well as swine flu virus specific isothermal gene amplification assay for early clinical diagnosis of H1N1 human patients.
Claims8
A process for detecting pandemic Swine flu virus H1N1 type in a sample, said process comprises providing the sample, mixing the sample with the oligonucleotides as set forth in SEQ ID NO:
- 1 to SEQ ID NO:
- 8 and buffer comprising reagents used for RNA amplification to obtain a reaction mixture, incubating said reaction mixture at a temperature of about 60° C. to about 65° C. for a period of about 15 minutes to about 45 minutes and detecting amplified product, wherein presence of the amplified products confirms presence of the virus.
A process of detecting pandemic Swine Flu Virus H1N1 type in a sample, wherein said process comprises providing the sample, obtaining cDNA from the sample using oligonucleotides as set forth in SEQ ID NO:
- 1 and SEQ ID NO:
- 2, mixing the cDNA obtained with the oligonucleotides as set forth in SEQ ID NO:
- 3 to SEQ ID NO:
- 8 and buffer comprising reagents used for DNA amplification to obtain a reaction mixture, incubating said reaction mixture at a temperature of about 60° C. to about 65° C. for a period of about 15 minutes to about 45 minutes and detecting amplified product, wherein presence of the amplified products confirms presence of the virus.
A primer set for detection of pandemic Swine Flu Virus H1N1 type in a sample comprising:
- (a) a forward outer primer (F3) comprising the oligonucleotide of SEQ ID NO:1 and
- (b) a backward outer primer (B3) comprising the oligonucleotide of SEQ ID NO:2a forward inner primer (FIP) comprising the oligonucleotide of SEQ ID NO:3, a linker and the oligonucleotide of SEQ ID NO:4;
- (c) a backward inner primer (BIP) comprising the oligonucleotide of SEQ ID NO:5, a linker and the oligonucleotide of SEQ ID NO:6;
- (d) a forward loop primer (FLP) comprising the oligonucleotide of SEQ ID NO:5;
- and
- (e) a backward loop primer (BLP) comprising the oligonucleotide of SEQ ID NO:
- 8.
Dependencies are inferred from the claim text.