Application of loop mediated isothermal amplification in the detection of bovine viral diarrhea

ZHOU Xian, CHEN Wenqin, WANG Wenxi, SUN Zhiwu, ZHANG Miaomiao, GUO Nini, SONG Xianrong, WU Xiuzhu

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Animals Breeding and Feed ›› 2025, Vol. 24 ›› Issue (05) : 41-46. DOI: 10.13300/j.cnki.cn42-1648/s.2025.05.011

Application of loop mediated isothermal amplification in the detection of bovine viral diarrhea

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Abstract

Objectives A rapid and sensitive method for detecting bovine viral diarrhea virus (BVDV) based on the loop-mediated isothermal amplification (LAMP) technology was developed to provide reference for the diagnosis of bovine viral diarrhea. Methods Multiple sets of primers and probes were designed based on the conserved sequence of 5'-UTR gene of BVDV in GenBank, and a set of primers and probes with the best effect of amplification were screened. A method for detecting BVDV with LAMP was established by optimizing the reaction system and annealing temperature. The sensitivity and specificity of the method established were evaluated, and the method was used to detect clinical samples. Results The LAMP method established had high specificity, producing amplification signals only for BVDV and showing no cross-reaction with common bovine pathogens including foot-and-mouth disease and Salmonella. Its sensitivity of detection verified by gradient dilution of recombinant plasmids was 102 copies/µ L, 10 times higher than that of conventional PCR. The results of detecting clinical samples were completely consistent with that of PCR method, with a concordance rate of 100%. Conclusions The established method of detecting BVDV with LAMP has high sensitivity and strong specificity. It will provide technical support for the clinical diagnosis and treatment of bovine viral diarrhea.

Key words

bovine viral diarrhea virus (BVDV) / loop-mediated isothermal amplification (LAMP) / rapid detection / clinical diagnosis and treatment / application

CLC number

S858.23

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ZHOU Xian , CHEN Wenqin , WANG Wenxi , et al . Application of loop mediated isothermal amplification in the detection of bovine viral diarrhea. Animals Breeding and Feed. 2025, 24(05): 41-46 https://doi.org/10.13300/j.cnki.cn42-1648/s.2025.05.011

References

1
PANG F LONG Q Q WEI M.Immune evasion strategies of bovine viral diarrhea virus[J/OL].Frontiers in cellular and infection microbiology202313:1282526[2024-12-19].
2
BACHOFEN C, BOLLINGER B, PETERHANS E, et al. Diagnostic gap in bovine viral diarrhea virus serology during the periparturient period in cattle[J]. J Vet Diagn Invest,2013,25:655-661.
3
STORINO G Y, MECHLER-DREIBI M L, XAVIER E B, et al. Artificial insemination of gilts with bovine viral diarrhea virus-contaminated semen[J]. Can Vet J, 2021,62:59-61.
4
牟思洋,李焱,何小强,等.牛病毒性腹泻病毒流行和防控研究进展[J].养殖与饲料2024,23(10):90-94.
5
NOTOMI T, OKAYAMA H, MASUBUCHI H, et al. Loop-mediated isothermal amplification of DNA[J/OL].Nucleic acids research, 2000,28(12):E63[2024-12-19].
6
DOMESLE K J, YANG Q, HAMMACK T S, et al. Validation of a Salmonella loop-mediated isothermal amplification assay in animal food[J]. Int J Food Microbiol,2018, 264:63-76.
7
SILVA S J R, PARDEE K, BALASURIYA U B R,et al. Development and validation of a one-step reverse transcription loop-mediated isothermal amplification (RT-LAMP) for rapid detection of ZIKV in patient samples from Brazil[J/OL]. Sci Rep,2019,11: 4494[2024-12-19].
8
朱杰,杨才俊,祁明普,等.我国牛群中牛病毒性腹泻病毒流行的Meta分析[J].华中农业大学学报2023,42(2):48-62.
9
郑如雯,黄涛,吴道义 等.牛病毒性腹泻病毒LAMP-LFD检测方法的建立及初步应用[J].畜牧兽医学报,2023,54(11):4745-4753.
10
TAPONEN S, PYÖRÄLÄ S. Coagulase-negative staphylococci as cause of bovine mastitis:not so different from Staphylococcus aureus [J]. Vet Microbiol, 2009,134:29-36.
11
吴小萍,周敏,许建勋,等.牛病毒性腹泻的诊断及防控措施[J].畜牧业环境,2024(10):34-37.
12
李家伟,郭利,杨勇,等.牛病毒性腹泻病毒LAMP检测方法的建立与应用[J].中国畜牧兽医,2015,42(12):3111-3118.

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