Genetic characterization of bovine coronavirus in Vietnam

Shin, Jihye; Tark, Dongseob; Le, Van Phan; Choe, SeEun; Mi, Ra; Park, Gyunam; Cho, In‐Soo; Nga, Bui Thi To; Lan, Nguyen Thi; An, Dong-Jun

doi:10.1007/s11262-019-01647-1

Cited by 18 publications

(19 citation statements)

References 15 publications

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“…The expansion of BCoV to camel hosts further illustrates its ability to cross species and to infect other similar ungulate hosts. Recently, surveillance of BCoV has been expanded into regions of East Asia, including China (35), South Korea (36), and Vietnam (37), and into the Caribbean (Cuba) (38). The spike genes of BCoV strains from these regions were all phylogenetically clustered into C1.…”

Section: Discussionmentioning

confidence: 99%

Diversity of Dromedary Camel Coronavirus HKU23 in African Camels Revealed Multiple Recombination Events among Closely Related Betacoronaviruses of the Subgenus Embecovirus

Chu

Miguel

et al. 2019

J Virol

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Genetic recombination has frequently been observed in coronaviruses. Here, we sequenced multiple complete genomes of dromedary camel coronavirus HKU23 (DcCoV-HKU23) from Nigeria, Morocco, and Ethiopia and identified several genomic positions indicative of cross-species virus recombination events among other betacoronaviruses of the subgenus Embecovirus (clade A beta-CoVs). Recombinant fragments of a rabbit coronavirus (RbCoV-HKU14) were identified at the hemagglutinin esterase gene position. Homolog fragments of a rodent CoV were also observed at 8.9-kDa open reading frame 4a at the 3= end of the spike gene. The patterns of recombination differed geographically across the African region, highlighting a mosaic structure of DcCoV-HKU23 genomes circulating in dromedaries. Our results highlighted active recombination of coronaviruses circulating in dromedaries and are also relevant to the emergence and evolution of other betacoronaviruses, including Middle East respiratory syndrome coronavirus (MERS-CoV).IMPORTANCE Genetic recombination is often demonstrated in coronaviruses and can result in host range expansion or alteration in tissue tropism. Here, we showed interspecies events of recombination of an endemic dromedary camel coronavirus, HKU23, with other clade A betacoronaviruses. Our results supported the possibility that the zoonotic pathogen MERS-CoV, which also cocirculates in the same camel species, may have undergone similar recombination events facilitating its emergence or may do so in its future evolution.

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Section: Discussionmentioning

confidence: 99%

Diversity of Dromedary Camel Coronavirus HKU23 in African Camels Revealed Multiple Recombination Events among Closely Related Betacoronaviruses of the Subgenus Embecovirus

Chu

Miguel

et al. 2019

J Virol

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“…Coronaviruses are enveloped RNA virus, spherical to pleomorphic in shape, with 20 nm spikes on the envelope. A member of the coronavirus family, within the beta A subtype [1,2], Bovine coronavirus (BCoV), is a respiratory and enteric pathogen that infects cattle and wild ruminant species. Like several other variations of coronaviruses, BCoV has additional 5 nm secondary spikes [2,3].…”

Section: Introductionmentioning

confidence: 99%

“…Like several other variations of coronaviruses, BCoV has additional 5 nm secondary spikes [2,3]. The larger spike interacts with sialic acid within receptors on the target cell, while the smaller spike contains hemagglutinin-esterase (HE), another protein critical in attachment to target cells [1,4]. The virus results in three distinct symptoms: calf diarrhea, winter dysentery with hemorrhagic diarrhea in adults, and respiratory infection in cattle of all ages [4,5].…”

Section: Introductionmentioning

confidence: 99%

“…These symptoms are devastating for the agricultural industry, with impacts including the cost of treatment and prophylaxis, increased vulnerability to additional diseases, increased mortality rates, long-term lowered growth rates, and lowered milk production [8,9]. Widespread BCoV infections can have disastrous effects, especially in countries like Uruguay or Vietnam, where cattle production is a primary source of income and the overall BCoV detection rate is 7.8% [7] and 33.3% [1], respectively. Current diagnostic method involves sampling feces and intestinal contents and performing reverse transcription-polymerase chain reactions (RT-PCR) to assess for BCoV [7][8][9].…”

Section: Introductionmentioning

confidence: 99%

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Dissecting Transcription Factor-Target Interaction in Bovine Coronavirus Infection

et al. 2020

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Coronaviruses are RNA viruses that cause significant disease within many species, including cattle. Bovine coronavirus (BCoV) infects cattle and wild ruminants, both as a respiratory and enteric pathogen, and possesses a significant economic threat to the cattle industry. Transcription factors are proteins that activate or inhibit transcription through DNA binding and have become new targets for disease therapies. This study utilized in silico tools to identify potential transcription factors that can serve as biomarkers for regulation of BCoV pathogenesis in cattle, both for testing and treatment. A total of 11 genes were identified as significantly expressed during BCoV infection through literature searches and functional analyses. Eleven transcription factors were predicted to target those genes (AREB6, YY1, LMO2, C-Rel, NKX2-5, E47, RORAlpha1, HLF, E4BP4, ARNT, CREB). Function, network, and phylogenetic analyses established the significance of many transcription factors within the immune response. This study establishes new information on the transcription factors and genes related to host-pathogen interactome in BCoV infection, particularly transcription factors YY1, AREB6, LMO2, and NKX2, which appear to have strong potential as diagnostic markers, and YY1 as a potential target for drug therapies.

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“…Bovine coronavirus (BCoV) is an important pathogen of NCD worldwide. Coronaviruses contains a positive sense single stranded RNA genome, coding for five major structural proteins: including the nucleocapsid (N), spike (S) and non-structural proteins [18,19,23].…”

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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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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Genetic characterization of bovine coronavirus in Vietnam

Cited by 18 publications

References 15 publications

Diversity of Dromedary Camel Coronavirus HKU23 in African Camels Revealed Multiple Recombination Events among Closely Related Betacoronaviruses of the Subgenus Embecovirus

Diversity of Dromedary Camel Coronavirus HKU23 in African Camels Revealed Multiple Recombination Events among Closely Related Betacoronaviruses of the Subgenus Embecovirus

Dissecting Transcription Factor-Target Interaction in Bovine Coronavirus Infection

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

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