New set of oligonucleotide primers comprising a forward primer and reverse primer, used in assay for in-vitro detection of Salmonella enterica in a sample, preferably food sample
	DUTT S A; MADAN R U; NANDKISHOR M C; VIJAY M; BABU K H; KINGSTON J J; MARY I A
	2023-08-11
专利权人	DRDO (DRDO-Non-standard)
申请日期	2023-08-11
专利号	IN202311053994-A
成果简介	NOVELTY - Set of oligonucleotide primers is new, where set of oligonucleotide primers comprises a forward primer comprising 18 nucleotide sequence (SEQ ID NO: 1), and a reverse primer comprising 19 nucleotide sequence (SEQ ID NO: 2), given in the specification. USE - Set of oligonucleotide primers used in assay for in-vitro detection of Salmonella enterica in a sample, preferably food sample. ADVANTAGE - The set of oligonucleotide primers has greater reproducibility and reduced chances of carry-over contamination, and shows 100% specificity and sensitivity of detection of Salmonella enterica in a sample, and no amplification in non-Salmonella strains. DETAILED DESCRIPTION - Set of oligonucleotide primers is new, where set of oligonucleotide primers comprises a forward primer comprising 18 nucleotide sequence caRcgctggggaaatgac (SEQ ID NO: 1), and a reverse primer comprising 19 nucleotide sequence gatcRtctcgcRYcaggtg (SEQ ID NO: 2). INDEPENDENT CLAIMS are included for: Probe, which comprises a fluorophore and a quencher linked to the probe, where the probe comprises 16 nucleic acid sequence taccgtcaaatacgca (SEQ ID NO: 13), given in the specification; Method for in-vitro detection of Salmonella enterica in a sample, which involves: (1) obtaining DNA from a sample; (2) obtaining a set of oligonucleotide primer; (3) obtaining a probe; (4) contacting the DNA of step (1) with the set of oligonucleotide primer of step (2), probe of step (3), a premix, ROX reference dye, and solvent to obtain a reaction mixture; (5) incubating the reaction mixture of step (4) at a thermal cycling condition of 40 cycles with an initial denaturation step carried out at a temperature of 95℃ for 20 seconds, followed by denaturation step carried out at a temperature of 95℃ for 3 seconds, and annealing and extension step carried out a temperature in the range of 55-62℃ for 35 seconds to obtain an amplified mixture; and (6) detecting for the presence or absence of an amplified target region in the amplified mixture, where the presence of the amplified target region in the amplified mixture indicates the presence of Salmonella enterica in the sample; Method for detection of Salmonella pathogen in a food sample, which involves: (1) spiking a food sample with a bacterium having 103-100 colony forming units (CFG)/g of the food; (2) homogenizing the food sample of step (1) in a shaking incubator at a temperature in the range of 35℃ to 40℃ at a speed in the range of 150-170 revolution/minute (rpm) to obtain an enriched mixture; (3) drawing samples in quadruplicates from the enriched mixture; and (4) performing a real-time PCR method using the set of oligonucleotide primer, and probe to detect the presence of Salmonella enterica in the food sample; and Assay for detecting the presence of Salmonella enterica in a sample, which comprises a set of oligonucleotide primer, probe, premix, ROX reference dye, and solvent that is water.
IPC 分类号	C07K-016/12 ; C12N-001/20 ; C12Q-001/689 ; C12Q-001/70 ; G01N-033/569
国家	印度
专业领域	医药卫生
语种	英语
成果类型	专利
文献类型	科技成果
条目标识符	http://119.78.100.226:8889/handle/3KE4DYBR/20872
专题	中国科学院新疆生态与地理研究所
作者单位	DRDO (DRDO-Non-standard)
推荐引用方式 GB/T 7714	DUTT S A,MADAN R U,NANDKISHOR M C,et al. New set of oligonucleotide primers comprising a forward primer and reverse primer, used in assay for in-vitro detection of Salmonella enterica in a sample, preferably food sample. IN202311053994-A[P]. 2023.

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