• Clinical and Experimental Pediatrics
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• v.56 no.9
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• pp.383-388
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• 2013

Purpose: Viral etiology is common in cases of children with acute diarrhea, and antibiotic therapy is usually not required. Therefore, it is important to determine the distribution of common viruses among children hospitalized with acute diarrhea. Methods: We included 186 children who suffered from acute diarrhea and were hospitalized at the Wonkwang University Hospital Pediatric ward from December 1, 2010 to June 30, 2011 in this study. Stool samples were collected and multiplex reverse transcriptase polymerase chain reaction (multiplex RT-PCR) was used to simultaneously determine the viral etiology such as rotavirus, norovirus, astrovirus, or adenovirus. Results: Causative viruses were detected in 72 of the 186 cases (38.7%). The mean age of the virus-positive cases was 1 year and 9 months (range, 1 month to 11 years). Rotavirus was detected in 50/186 (26.9%); norovirus, in 18/186 (9.7%); and astrovirus, in 3/186 cases (1.6%). Adenovirus was not detected in any of the cases. Proportions of norovirus genogroups I and II were 21.1% and 78.9%, respectively. Four of the 51 rotavirus-positive cases (7.8%) had received rotavirus vaccination at least once. The mean duration of diarrhea was 2.8 days (range, 1 to 10 days) and vomiting occurred in 39 of the 72 cases (54.2%). Conclusion: Viral etiology was confirmed in about one-third of the children with acute diarrhea, and the most common viral agent was rotavirus, followed by norovirus.

• Microbiology and Biotechnology Letters
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• v.40 no.4
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• pp.339-347
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• 2012

Viral safety is an important prerequisite for clinical preparations of mammalian cell culture-derived biopharmaceuticals, because numerous adventitious viruses have been contaminated during the manufacturing process. In particular, Chinese hamster ovary (CHO) cells are highly susceptible to several RNA viruses including reovirus (Reo), bovine viral diarrhea virus (BVDV), and bovine parainfluenza virus (BPIV) and there have been reports of such viral contaminations. Therefore, viral detection during the CHO cell process is necessary to ensure the safety of biopharmaceuticals against viruses. In this study, a multiplex reverse transcription (RT)-PCR assay was developed and subsequently evaluated for its effectiveness as a means to simultaneously detect Reo, BVDV, and BPIV during the manufacture of cell culture-derived biopharmaceuticals. Specific primers for Reo, BVDV, and BPIV were selected, and a multiplex RT-PCR was optimized. The sensitivity of the assay for simultaneous amplification of all viral target RNAs was $7.76{\times}10^2\;TCID_{50}/ml$ for Reo, $7.44{\times}10^1\;TCID_{50}/ml$ for BVDV, and $6.75{\times}10^1\;TCID_{50}/ml$ for BPIV. The multiplex RT-PCR was proven to be very specific to Reo, BVDV, and BPIV and was subsequently applied to the validation of CHO cells artificially infected with each virus. It could detect each viral RNA from CHO cells as well as culture supernatants. Therefore, it was concluded that the multiplex RT-PCR assay can be applied to detection of the adventitious viruses during the manufacture of cell culture-derived biopharmaceuticals.

• Journal of Veterinary Clinics
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• v.26 no.6
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• pp.578-581
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• 2009

A 25-day-old, Korean indigenous calf was presented with a 10 days history of respiratory disorders and bloody diarrhea, and died. This calf was extremely unthrifty compared to others and had evidence of chronic diarrhea based on matting of feces in the hair of the tail and perineum. Ecchymotic hemorrhages were observed on multiple organs at necropsy. Bovine viral diarrhea virus (BVDV) infection was identified by RT-PCR. The phylogenetic analysis showed that this case belonged to BVDV-2a subgroup and was related to highly pathogenic USA isolate 890 (U18059). This case provided evidence for circulation of BVDV-2 in Republic of Korea. The occurrence of BVDV-2 was also reconfirmed.

• Korean Journal of Veterinary Research
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• v.60 no.3
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• pp.183-186
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• 2020

This paper describes the development of neurological signs of two prematurely born calves four days after birth. The pathological examination results indicated fibrinopurulent polyserositis, including meningoencephalitis with suppurative bronchopneumonia. Bovine viral diarrhea virus subtype 2a was detected in most of the internal organs, and the bacterial colonies cultured from the samples were identified as Klebsiella (K.) pneumoniae. Molecular analysis via multilocus sequence typing identified a different K. pneumoniae isolate in each calf-type 14 in calf A and type 65 in calf B. This is the first report identifying K. pneumoniae sequence types 14 and 65 in cattle.

• Journal of Veterinary Clinics
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• v.24 no.2
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• pp.150-153
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• 2007

The aim of this study was to investigate the prevalence of bovine viral diarrhea virus (BVDV) infection in Chonbuk province. Blood samples were taken from 92 korean calves to determined their serological status against BVDV, Capture enzyme-linked immunosorbent assay (ELISA) was used to test for antigen. The number of seropositive calves ranged from 3.3% to 12.9%. Antigens against BVDV were detected in 3.3% of healthy calves, 6.4% of digestive symptom calves, 12.9% of respiratory symptom calves, respectively. Sex and age of calves had no significant differences on the prevalence of BVDV. The results indicate that transmission of BVDV may have become exposed as a result of contact with acute infected or persistently infected cattle.

• Journal of Veterinary Clinics
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• v.24 no.2
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• pp.169-171
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• 2007

The aim of this study was to detect bovine viral diarrhea virus (BVDV) from calves in Chonbuk province. Blood samples were taken from ninety-two dairy calves. Capture enzyme-linked immunosorbent assay (ELISA) was used to detect BVDV. BVDV were detected in eight out of ninety-two (8.6%) dairy calves. BVDV were detected in one of twenty five of female calves and one of twenty three of male calves of 4 months old, whereas in the 5 months age group, BVDV were detected in low of twenty three of female calves and two of twenty one of male calves. There were no significant differences (p>0.05) in the detection rate of BVDV on the basis of sex. On the other hand, ages of calves had significant differences (p<0.05) on the prevalence of BVDV.

• Journal of Practical Agriculture & Fisheries Research
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• v.5 no.1
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• pp.57-63
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• 2003

The bovine rotavirus(BRV), bovine coronavirus(BCV) and bovine viral diarrhea virus(BVDV) are main viruses of bovine viral diarrhea disease. These viruses could be rapidly amplified by the reverse transcriptase polymerase chain reaction(RT-PCR). This study was conducted to develop rapid and accurate diagnostic methods of these viral diseases by multiplex RT-PCR. Specific primers were designed based on the sequences reported by Chang KO et. al. (1997) and Schroeder BA, et. al. (1990), RNA were prepared from the cultured viruses, first-stranded DNAs were synthesised by reverse transcriptase. PCR were conducted to amplify specific regions of the viruses by multiplex. Three bands such as 1,062bp for BRV, 458bp for BCV, and 300bp for BVDV were successfully produced by multiplex RT-PCR. In conclusion, this result suggested that these viruses could be diagnosed rapidly and accurately by multiplex RT-PCR.

• Journal of Veterinary Clinics
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• v.23 no.3
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• pp.300-307
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• 2006

In the present study, methods of the reverse transcription-polymerase chain reaction(RT-PCR) were evaluated for the rapid detection and differentiation of transmissible gastroenteritis virus(TGEV), porcine epidemic diarrhea virus(PEDV) and rotavirus in piglets suffering from diarrhea. For the purposes, the PCR conditions were first confirmed for the amplification of VP7 gene of rotavirus and N gene of TGEV and PEDV using each specific primers and their annealing temperature. Multiplex RT-PCR methods were further determined to distinguish these viral infections and the results are as follows. For the specific amplification of these viral genes, the reliable PCR condition was determined as 30 cycles of reaction consisting each 1 min of denature at $94^{\circ}C$, annealing at $42^{\circ}C$ and polymerization at $72^{\circ}C$ with 1.0 mM $MgCl_2$. It was able to differentiate these viral infections in the intestines and feces of piglets suffering from diarrhea by duplex PCR for TGEV and PEDV and single PCR for rotavirus with a primer-annealing temperature of $42^{\circ}C$. When the multiplex RT-PCR were undertaken for the field samples, 17 cases of PEDV and 5 cases of rotavirus infections were differential diagnosed in a total of 92 samples of intestines and feces of the piglets with diarrhea.

• Korean Journal of Veterinary Research
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• v.39 no.4
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• pp.743-750
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• 1999

Bovine viral diarrhea viruses (BVDVs) were isolated from cattle with respiratory and diarrhea signs as well as persistently infected cattle. These isolates were analysed serologically to characterize serogroups and to compare serological relationship with reference viruses of type I and II. Most isolates from calf diarrheal cases and persistently infected individuals showed a significant difference in cross-neutralization test with the viruses isolated from nasal discharges showing severe respiratory signs. Serologically most of the commercial vaccine strains could be classified into classical BVDV (type I) such as NADL strain. This serological difference among BVDV isolates suggested the need for new vaccines to protect cattle from both respiratory and enteric BVDV infections in field. The immunogenicity of BVDVs which showed a good propagation capability in MDBK cells and high rates of neutralizing activity (isolate : KD26-1, PHG, B5 and 95002) against all viruses used in this study, was confirmed in guinea pig when treated as single or combined groups.

• Korean Journal of Veterinary Service
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• v.20 no.4
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• pp.331-338
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• 1997

The purpose of this study was to establish early diagnosis for bovine virus diarrhea-mucosal disease(BVD-MD) Two Holstein among 22 breeding cows were shown ulcer in the mouth and watery diarrhea. Diarrheal feces and ulcerous lesion of the mouth from 2 cows were sampled for detection of viral antigen. BVD virus was isolated by inoculation of the samples to MDBK cells, and the cytopathic effects were observed in cultured MDBK cells which inoculated with virus isolates from the feces. Viral antigens were detected in the feces and ulceruous lesion by immunogold staining. The serum neutralization titers were shown 1 : 64 or greater in 8 blood samples by using BVD virus (NADL strain). By the RT-PCR, using reverse primer 5'-ACTCCATGTGCCATGTACAG-3', forward primer 5'-ACTCCATGTGCCATGTACAG-3', 285 base pair band specific to BVD virus was detected. In conclusions, the results of above tests which executed using the diarrheal feces and ulcerous lesion of the mouth and the isolates were conformed as BVD virus.