Development and use of a reverse transcription insulated isothermal PCR assay for detection and characterization of bovine torovirus in yaks

Zhao, Long; Shao, Guoqing; Tang, Cheng; Yue, Hua

doi:10.1007/s00705-021-05047-5

Cited by 5 publications

(2 citation statements)

References 38 publications

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“…There are several rapid diagnostic assays currently available for the detection of yak-derived components [16]. However, most of these assays are complicated, costly, and time-consuming, making them difficult to implement at the grassroots level and unsuitable for on-site rapid testing.…”

Section: Discussionmentioning

confidence: 99%

The Identification of Yak Meat Using Loop-Mediated Isothermal Amplification Method Coupled with Hydroxy Naphthol Blue for the Prevention of Food Fraud

Zhao,

Tan,

Wang

et al. 2024

Journal of Food Processing and Preservation

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Objective. Yak is found in the Qinghai-Tibet Plateau and represents a meat of high nutritional value and good flavor. However, the production of yak is limited, and yak meat adulteration is a growing concern in the marketplace. To protect consumer rights and prevent unfair competition, it is necessary to use an efficient assay to identify the species of yak meat rapidly and accurately being sold. Methods. Loop-mediated isothermal amplification (LAMP) combined with hydroxy naphthol blue (HNB) was used to identify potential adulterants. The specificity and sensitivity tests of yak-derived components were carried out to achieve the monitoring of yak-derived components. Results. The optimal color development was achieved with an external primer-to-internal primer ratio of 400 nmol/L : 1200 nmol/L, 1.5 mmol/L dNTP, and 0.32 U/μL Bst DNA polymerase with 5 mmol/L MgSO4 at 62°C amplification temperature. The detection sensitivity of LAMP-HNB for yak-derived DNA was up to 1 pg/μL. Conclusion. The LAMP-HNB assays provided a valuable tool for the identification of yak gene from adulterated meat. This further enabled the LAMP-HNB assay to be applicable in the identification of other meat products.

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Section: Discussionmentioning

confidence: 99%

The Identification of Yak Meat Using Loop-Mediated Isothermal Amplification Method Coupled with Hydroxy Naphthol Blue for the Prevention of Food Fraud

Zhao,

Tan,

Wang

et al. 2024

Journal of Food Processing and Preservation

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“…Specifically, a high prevalence of BToV infection has been observed among 3 week-old calves with diarrhea (Hoet et al, 2003). Currently, BToV has been reported in 17 countries worldwide, which has attracted increasing attention (Zhao et al, 2021).…”

Section: Introductionmentioning

confidence: 99%

A multiplex real-time fluorescence-based quantitative PCR assay for calf diarrhea viruses

Meng,

Chen,

Jiang

et al. 2024

Front. Microbiol.

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IntroductionCalf diarrhea is a significant condition that has a strong effect on the cattle industry, resulting in huge economic losses annually. Bovine torovirus (BToV), bovine enterovirus (BEV), bovine norovirus (BNoV), bovine coronavirus (BCoV), bovine rotavirus (BRV), and bovine viral diarrhea virus (BVDV) are key pathogens that have been implicated in calf diarrhea. Among these viruses, there remains limited research on BToV, BEV, and BNoV, with no available vaccines or drugs for their prevention and control. Although commercial vaccines exist for BCoV, BRV, and BVDV, the prevalence of these diseases remains high.MethodsTo address this issue, we developed a multiplex real-time fluorescence quantitative PCR method for detecting BToV, BEV, BNoV, BCoV, BRV, and BVDV. This method can be used to effectively monitor the prevalence of these six viruses and serve as a reference for future prevention and control strategies. In this study, we specifically designed primers and probes for the BNoV Rdrp, BEV 5′UTR, BToV M, BCoV N, BRV NSP5, and BVDV 5′UTR genes.ResultsThis method was determined to be efficient, stable, and sensitive. The lowest detectable levels of plasmids for BNoV, BEV, BToV, BRV, BCoV, and BVDV were 1.91 copies/μL, 96.0 copies/μL, 12.8 copies/μL, 16.4 copies/μL, 18.2 copies/μL, and 65.3 copies/μL, respectively. Moreover, the coefficients of variation for all six detection methods were < 3%; they also exhibited a strong linear relationship (R2 ≥ 0.98), and an amplification efficiency of 90%−110%. A total of 295 fecal and anal swabs were collected from calves with diarrhea in Guangdong, China. The positive rates for BToV, BEV, BNoV, BCoV, BR, and BVDV were determined to be 0.34% (1/295), 6.10% (18/295), 0.68% (2/295), 1.36% (4/295), 10.85% (32/295), and 2.03% (6/295), respectively. Notably, BEV and BRV exhibited the highest prevalence.DiscussionAdditionally, this study identified the occurrence of BToV and BNoV in Guangdong for the first time. In summary, this study successfully established an effective method for detecting several important bovine viruses; ultimately, this holds strong implications for the future development of the cattle industry.

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torovirus infections

Behboudi

2023

CABI Compendium

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Development and use of a reverse transcription insulated isothermal PCR assay for detection and characterization of bovine torovirus in yaks

Cited by 5 publications

References 38 publications

The Identification of Yak Meat Using Loop-Mediated Isothermal Amplification Method Coupled with Hydroxy Naphthol Blue for the Prevention of Food Fraud

The Identification of Yak Meat Using Loop-Mediated Isothermal Amplification Method Coupled with Hydroxy Naphthol Blue for the Prevention of Food Fraud

A multiplex real-time fluorescence-based quantitative PCR assay for calf diarrhea viruses

torovirus infections

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