+ Site Statistics
+ Search Articles
+ PDF Full Text Service
How our service works
Request PDF Full Text
+ Follow Us
Follow on Facebook
Follow on Twitter
Follow on LinkedIn
+ Subscribe to Site Feeds
Most Shared
PDF Full Text
+ Translate
+ Recently Requested

Cross-protection studies between respiratory and calf diarrhea and winter dysentery coronavirus strains in calves and RT-PCR and nested PCR for their detection



Cross-protection studies between respiratory and calf diarrhea and winter dysentery coronavirus strains in calves and RT-PCR and nested PCR for their detection



Archives of Virology 146(12): 2401-2419



A 1-step RT-PCR assay, targeting a 730 bp fragment of the nucleocapsid (N) gene of bovine coronavirus (BCV), and a nested PCR assay, targeting a 407 bp fragment of the N gene, were developed to detect BCV in nasal swab and fecal samples of calves experimentally exposed to BCV. Both 1-step RT-PCR and nested PCR recognized cell culture passaged isolates of 10 bovine respiratory coronavirus (BRCV), 5 calf diarrhea (CD) and 8 winter dysentery (WD) strains of BCV, but not transmissible gastroenteritis coronavirus or bovine rotavirus. The sensitivity of the 1-step RT-PCR and nested PCR was compared to that of an antigen-capture ELISA. The lowest detection limit of the 1-step RT-PCR and nested PCR as determined by using tenfold serial dilutions of the BRCV 255 and 440 strains in BCV negative nasal swab suspensions from preexposure gnotobiotic calves was 2 x 10(4) and 2 x 10(2) TCID50/0.1 ml for each strain, respectively. The lowest detection limit of the antigen-capture ELISA as determined by using the same serially diluted samples was 1 x 10(6) TCID50/0.1 ml for each strain. Therefore, the 1-step RT-PCR and nested PCR assays were 50 and 5000 times, respectively more sensitive than the antigen-capture ELISA to detect BRCV in nasal swab suspensions. To investigate in vivo cross-protection between the BRCV and CD or WD strains of BCV and to detect nasal and fecal shedding of BCV using the 1-step RT-PCR, nested PCR and antigen-capture ELISA, 6 colostrum-deprived and two gnotobiotic calves were inoculated with a BRCV, a CD or a WD strain of BCV and then challenged 3-4 weeks later with either BRCV, CD or WD strains of BCV. All calves developed diarrhea after inoculation and BCV antigen (ELISA) or RNA (RT-PCR) was detected in the diarrheic fecal samples or the corresponding nasal swab samples. In addition, low amounts of BCV were also detected only by nested PCR in the fecal and nasal swab samples before and after diarrhea. No respiratory clinical signs were observed during the entire experimental period, but elevated rectal temperatures were detected during diarrhea in the BCV-inoculated calves. All calves recovered from infection with the BRCV, CD, or WD strains of BCV were protected from BCV-associated diarrhea after challenge exposure with either a heterologous or homologous strain of BCV. However, all calves challenged with heterologous BCV strains showed subclinical BCV infection evident by detection of nasal and fecal shedding of BCV RNA detected only by nested PCR. Such results confirm field and experimental data documenting reinfection of the respiratory and enteric tracts of cattle, suggesting that, in closed herds, respiratory or enteric tract reinfections may constitute a source of BCV transmissible to cows (WD) or neonatal or feedlot calves. In addition, the present 1-step RT-PCR and nested PCR assays were highly sensitive to detect BCV in nasal swab and fecal specimens. Therefore, these assays should be useful to diagnose BCV infections in calves and adult cows.

Please choose payment method:






(PDF emailed within 1 workday: $29.90)

Accession: 045674500

Download citation: RISBibTeXText

PMID: 11811688


Related references

Infection and cross-protection studies of winter dysentery and calf diarrhea bovine coronavirus strains in colostrum-deprived and gnotobiotic calves. American Journal of Veterinary Research 57(1): 48-53, 1996

Detection of bovine coronavirus and type A rotavirus in neonatal calf diarrhea and winter dysentery of cattle in Quebec: evaluation of three diagnostic methods. Canadian Veterinary Journal 35(3): 163-169, 1994

Dual enteric and respiratory tropisms of winter dysentery bovine coronavirus in calves. Archives of Virology 152(10): 1885-1900, 2007

Calf diarrhea coronavirus: studies for the detection and isolation of the virus. Dissertation Abstracts International, B 43(11): 3501, 1983

Coronavirus-associated diarrhea (winter dysentery) in adult cattle. Canadian Veterinary Journal 30(10): 825-827, 1989

Protection studies on winter dysentery caused by bovine coronavirus in cattle using antigens prepared from infected cell lysates. Canadian Journal of Veterinary Research 64(2): 138-140, 2000

Comparative diagnostic studies of fecal samples (EM, virus isolation, immunological method) for the detection of bovine Coronavirus in calves with diarrhea. 1990

Canine coronavirus detection in feces from diarrhea and healthy dogs by nested-PCR. Virologica Sinica 20(1): 41-45, 2005

Comparison of bovine coronavirus isolates associated with neonatal calf diarrhoea and winter dysentery in adult dairy cattle in Quebec. Journal of general virology 76(5): 1263-1270, 1995

Genomic and antigenic variations of the HE glycoprotein of bovine coronaviruses associated with neonatal calf diarrhea and winter dysentery. Archives of Virology 146(6): 1219-1230, 2001

Bovine epizootic diarrhea resembling winter dysentery caused by bovine coronavirus. JARQ Japan Agricultural Research Quarterly 17(3): 185-190, 1983

Cross-protection against a human enteric coronavirus and a virulent bovine enteric coronavirus in gnotobiotic calves. Journal of Virology 80(24): 12350-6, 2006

Antigenic and biological comparisons of bovine coronaviruses derived from neonatal calf diarrhea and winter dysentery of adult cattle. Archives of virology40(7): 1303-1311, 1995

Winter dysentery in adult dairy cattle: detection of coronavirus in the faeces. Veterinary Record 123(11): 300-301, 1988

Detection of neonatal calf diarrhea virus, infant reovirus-like diarrhea virus, and a coronavirus using the fluorescent virus precipitin test. Journal of Clinical Microbiology 3(3): 376-377, 1976