First detection of bovine noroviruses and detection of bovine coronavirus in Australian dairy cattle

Symes, S.J.; Allen, Joanne L.; Woodward, KL; Bailey, Kirsten E; Gilkerson, James R.; Browning, Glenn F.

doi:10.1111/avj.12695

Cited by 9 publications

(10 citation statements)

References 36 publications

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“…The outcomes demonstrated that the sensitivity and specificity of qRT-PCR were greater than universal PCR. Bovine coronavirus (BCoV) was first identified during an outbreak of diarrhea among neonatal calves in the 1970s [15,21]. Earlier report demonstrated the existence of persistent infection of BCoV in cattle [11].…”

Section: Resultsmentioning

confidence: 99%

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Specific Detection of Bovine Coronavirus N Protein with TaqMan Probe qRT-PCR

Geng

Gong

et al. 2019

Acta Scientiae. Vet.

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Background: Bovine Coronavirus (BCoV) can cause acute diarrhea in newborn calves and adult cattle. BCoV infection may cause losses to production by reduced weight gain, reduced milk yield. Several methods have been applied to detect and diagnose BCoV. However, each assay has its deficiency. Currently, real-time quantitative PCR (qRT-PCR) has been utilized to identify and quantify many viral pathogens since it is a highly sensitive. However, the technical assay varies due to normalization control of the signal with an internal standard, typically a housekeeping gene. The main objective of the present study to establish a novel TaqMan probe real-time PCR (qRT-PCR) for detecting BCoV.Materials, Methods & Results: The present study was aimed to establish a novel TaqMan probe real-time PCR (qRT-PCR) for detecting bovine coronaviruses (BCoV), and also to develop a diagnostic protocol which simplifies sample collection and processing. One pair of specific primers, one pair of universal primers and a TaqMan probe were designed from the known sequences of conserved nucleocapsid (N) protein of BCoV. Reaction systems of TaqMan qRT-PCR were optimized including concentrations of the primers and probe as well as annealing temperatures. Prior to optimizing the assay, the recombinant plasmids of pMD18-T-BCoV-N were successfully constructed to make standard curves. The sensitivity, specificity and reproducibility were evaluated on the TaqMan qRT-PCR, respectively. A total of 321 feces specimens collected from diarrheic calves were detected with this assay. The results showed the optimized reaction conditions for qRT-PCR were 14.5 μM/L primers, 19.5 μM/L probes and 45.0°C annealing temperatures. The established TaqMan qRT-PCR assay could specially detect BCoV without detecting any other viruses. Its minimum detection limit was 4.72 × 101 copies/μL. However, universal PCR could detect only 4.72 × 103 copies/μL. Its sensitivity was 100-fold stronger than universal PCR. In conclusion, this TaqMan qRT-PCR had excellent specificity, sensitivity and stability with a 100-fold sensitivity stronger than universal PCR. Minimum detection limit was 4.72 × 101 copies/μL. This method was a cost-effective method to diagnose diarrhea and distinguish pathogens in dairy farms.Discussion: In this study, the authors developed a quantitative real-time PCR (qRT-PCR) in this study based on the TaqMan probe of BCoV. This TaqMan qRT-PCR assay selected and used one pair of specific primers (BCoV-qF/BCoV-qR) and a specific TaqMan probe (BCoV-probe) targeting the conserved nucleocapsid (N) gene. The specificity of primers and probes was validated with Primer-BLAST. The specificity of the qRT-PCR was confirmed by the negative control and other six viruses. The findings demonstrated that TaqMan qRT-PCR could only detect BCoV. This verified the qRT-PCR had an excellent specificity. It is obvious that this TaqMan qRT-PCR assay can detect only BCoV with stronger sensitivity and reproducibility than other real-time PCR methods. The sensitivity test indicated the minimum detection limit of the TaqMan qRT-PCR was 4.72 × 101copies/μL, or 47.2 copies/μL. Sensitivity of the TaqMan qRT-PCR assay was increased by 100-fold as compared to universal PCR with a good inter-assay and intra-assay reproducibility. Thereby, based on the high sensitivity of the assay of this qRT-PCR assay it may be a cost-effective method to diagnose BCoV infections and indentify the etiologic agents of diarrhea syndrome in the dairy farms.

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Section: Resultsmentioning

confidence: 99%

“…Several methods have been applied to detect and diagnose BCoV, such as viral culture, antigencapture ELISA, hemagglutination assay and PCR [4,21]. The universal PCR was reported for the detection of several animal and human viruses [5,17].…”

Section: Introductionmentioning

confidence: 99%

Specific Detection of Bovine Coronavirus N Protein with TaqMan Probe qRT-PCR

Geng

Gong

et al. 2019

Acta Scientiae. Vet.

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“…Recently, an updated classification of norovirus genogroups and genotypes was published (Chhabra et al., 2019); however, this update was focused on human norovirus. On the other hand, several studies have been recently performed in order to characterize norovirus circulating in cattle (Castells et al., 2020; Guo et al., 2018; Karayel‐Hacioglu & Alkan, 2019; Mohamed et al., 2017, 2018; Pourasgari et al., 2018; Ryu et al., 2020; Shi et al., 2019; Symes et al., 2018; Turan et al., 2018; Wang et al., 2019 ). A timely, exhaustive, and complete classification of GIII noroviruses is clearly needed, and in this work, we fill this need.…”

Section: Discussionmentioning

confidence: 99%

Evolutionary history and spatiotemporal dynamic of GIII norovirus: From emergence to classification in four genotypes

Castells

Cristina

Colina

2021

Transbounding Emerging Dis

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Noroviruses belong to a genetically diverse group of viruses infecting a wide range of mammalian host species, and those detected in cattle and sheep are classified within genogroup III (GIII). The current classification of norovirus in genogroups and genotypes is based on phylogenetic clustering and average distances within and between these phylogenetic clusters; however, the classification studies have been focused mainly on human norovirus, being GIII norovirus relegated. Due to the increasing number of studies on GIII norovirus, the need of an updated and extensive classification is evident. The aim of this study was to update the classification of norovirus within GIII, to describe the emergence of a circulating recombinant strain, and to reconstruct the evolutionary history of this genogroup. Two P‐types (GIII.P1–2) and four genotypes (GIII.1–4) were described. For the genogroup GIII, the evolutionary rate estimated was 2.78E‐3 s/s/y (95%HPD, 1.79E‐3 s/s/y–3.78E‐3 s/s/y), and the tMRCA was estimated around 1500 (95%HPD, 1247–1688). Despite the long history of this genogroup, the genotypes detected at present emerged in the last 100 years. Interestingly, most of the recombinant GIII.2P[1] strains detected worldwide were originated from a single recombination event and this recombinant strain was later dispersed through the world. Finally, our results indicate that a scenario of genotypes replacement through the time is highly probable.

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“…A study from Victoria describes the first isolation in Australia of bovine norovirus nucleotide sequences using rotavirus‐negative pooled faecal samples by PCR …”

Section: Production Animalsmentioning

confidence: 99%

In this issue – June 2018: Diversity of Haemophilus parasuis isolates from Australian piggeries · Detection of bovine noroviruses in Australian dairy cattle · Myocardial adenomatoid tumour in a calf · Hypertrophic cardiomyopathy in a horse · A history of veterinary education · Review of Q fever notifications in Victoria

Jackson¹

2018

Aust Veterinary J

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First detection of bovine noroviruses and detection of bovine coronavirus in Australian dairy cattle

Cited by 9 publications

References 36 publications

Specific Detection of Bovine Coronavirus N Protein with TaqMan Probe qRT-PCR

Specific Detection of Bovine Coronavirus N Protein with TaqMan Probe qRT-PCR

Evolutionary history and spatiotemporal dynamic of GIII norovirus: From emergence to classification in four genotypes

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