SYBR Green I RT-PCR Assay for Quantitative Detection of Salmonella in Irrigation Water

LV Xin; LIU Lanying; LI Yueren

doi:10.19303/j.issn.1008-0384.2023.08.013

Volume 38 Issue 8

Aug. 2023

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Article Navigation > Fujian Journal of Agricultural Sciences > 2023 > 38(8): 983-988

LV X, LIU L Y, LI Y R. SYBR Green I RT-PCR Assay for Quantitative Detection of Salmonella in Irrigation Water [J]. Fujian Journal of Agricultural Sciences，2023，38（8）：983−988 doi: 10.19303/j.issn.1008-0384.2023.08.013

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LV X, LIU L Y, LI Y R. SYBR Green I RT-PCR Assay for Quantitative Detection of Salmonella in Irrigation Water [J]. Fujian Journal of Agricultural Sciences，2023，38（8）：983−988 doi: 10.19303/j.issn.1008-0384.2023.08.013

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SYBR Green I RT-PCR Assay for Quantitative Detection of Salmonella in Irrigation Water

doi: 10.19303/j.issn.1008-0384.2023.08.013

LV Xin^{1, 2
,},
LIU Lanying^{1, 2},
LI Yueren^{1, 2
,
,}

1.
Institute of Quality Standards & Testing Technology for Agro-Products, Fujian Academy of Agricultural Sciences, Fuzhou, Fujian　350003, China
2.
Fujian Key Laboratory of Agro-products Quality & Safety, Fuzhou, Fujian　350003, China

Received Date: 2023-02-10
Rev Recd Date: 2023-05-22

Available Online: 2023-08-16

Publish Date: 2023-08-28

Abstract

Abstract

Objective A SYBR Green I-based quantitative RT-PCR (qPCR) method was developed for detecting salmonella in irrigation water. Method For the methodology development a pair of primers were designed based on the sequences of the invasion protein A gene (invA) of Salmonella. qPCR reaction conditions were optimized, the assay tested for specificity and sensitivity, and a standard curve of amplification constructed. Test results on a specimen of contaminated irrigation water using the qPCR assay were compared for detection accuracy with those obtained from the national standard method. Result The newly developed qPCR assay showed a minimum detection limit of 1×10⁻¹pg·μL⁻¹ and free of interference from genomic nucleic acids of non-target microbes. The constructed linear standard curve between 2×10⁰cfu·mL⁻¹ and 2×10⁵ cfu·mL⁻¹ had a high correlation coefficient of 0.999 6. The assay demonstrated same accuracy as did the national standard method in detecting salmonella in irrigation water. Conclusion The newly established qPCR assay could be adequately applied for salmonella detection in agriculture irrigation water.
- Quantitative RT-PCR,
- Irrigated water,
- Salmonella

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References(23)

References

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