• 미생물학회지
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• 제41권3호
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• pp.216-224
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• 2005

혈장분획제제 중 혈액응공인자제제와 일부 면역글로불린제제는 혈장에 존재하는 다양한 단백질로부터 유효한 단백성분만을 선택적으로 분리 정제하기 위해 크로마토그래피 방법을 사용하여 생산된다. 효율적인 세척(cleaning) 공정이 이루어지지 않는다면 크로마토그래피는 다양한 종류의 불순물뿐만 아니라 혈액 중 내재 또는 오염 가능성이 있는 위해인자가 오염될 가능성이 있다. 본 연구에서는 혈장분획제제 제조공정에 사용되는 크로마토그래피의 세척 공정에서 혈장유래 바이러스의 제거 및 불활화 공정의 검토 강화로 혈장분획제제의 안전성을 확보하기 위해 크로마토그래피 세척 검증 시스템을 구축하고자 하였다. 크로마토그래피 세척 공정 중 바이러스 제거 검증을 위해 혈장유래 바이러스 중 물리${\cdot}$화학적 처리에 가장 큰 저항성을 갖는 human parvovirus B19의 모델 바이러스의 porcine parvovirus(PPV)를 대상으로 real-time PCR 정량법을 확립하였다. PPV에 특이적인 primer를 선별하였으며 형광염료 SYBR Green I을 사용하여 PPV DNA를 정량하였다. 세포배양법에 의한 감염 역가와 비교한 결과 PCR 민감도는 1.5 $TCID_{50}/ml$이었다. 확립된 검증법의 신뢰성(reliability)을 보증하기 위해 실험법의 특이성(specificity), 재현성(reproducibility) 등을 검증하였다. 구축된 검증시스템을 thrombin 분리${\cdot}$정제를 위한 SP-Sepharose 양이온 크로마토그래피 공정과 factor VIII 분리${\cdot}$정제를 위한 Q-Sepharose 음이온 크로마토그래피 공정에 적용하여 크로마토그래피 세척 검증을 실시하고, 세척 검증 시스템의 적합성을 확인하였다.

• 대한수의학회지
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• 제36권4호
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• pp.895-902
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• 1996

산란계에 불활화 개 파보바이러스 백신을 근육내로 1주 간격으로 4회 접종하여 면역화시키고 최종 접종 2주후에 채란하여 $4^{\circ}C$에 보관하며 사용하였다. 난황항체는 5% HPMCP를 이용하여 분리하였고 0.5% HPMCP 용액은 lipid 침전에 매우 효과적이었으며 희석배수 10배에서 투명한 상층액을 나타내었다. 1차분리한 상층액의 단백질 농도는 2.5mg/ml이었고 최종 단백질 용액의 경우는 26.53mg/ml이었다. SDS-PAGE 전기영동상에서 분자량 60~70 KD 및 30~40 KD의 2 band가 나타났으며 non-reducing 전기영동에서는 닭 혈청 IgG와 같은 120~160 KD의 분자량을 보인 band가 각각의 분리용액에서 나타났다. 난황 항체의 개 파보바이러스에 대한 혈구응집억제반응 항체역가는 혈청의 역가에 비해 1주의 차이를 주며 증가했으며 난황 항체는 1:640에서 1:2560, 혈청은 1:640에서 1:5120을 나타내었다.

• 대한수의학회지
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• 제39권4호
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• pp.838-845
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• 1999

To determine the prognostic factors for survival of dogs infected with canine parvovirus, clinical and laboratory data of 35 dogs with clinical signs compatible with canine parvoviral enteritis admitted to the Veterinary Medical Teaching Hospital, Seoul National University during the period 1997-1998 were collected. Dogs were grouped by some major covariates, which can be considered as guides to the relative prognosis of dogs in the different subgroups. The Kaplan-Meier survival analysis and Weibull proportional hazard model were used to estimate overall survival, evaluate the comparability between groups, and identify potential prognostic factors. The overall survival rate for all dogs was 45.7% over the study period, and the Kaplan-Meier estimate of one week survival was 0.4989. Gender was the most favorable prognosis ; male dog (median, 6 days) had significantly higher risk of dying than female dog (median, 17 days ; p = 0.0023). In addition to gender, age was significantly associated with survival, with juvenile dogs less than 6-month-old having higher risk (p = 0.0359). Dogs that vaccinated with complete protocol (p = 0.0374) and those of having higher value of mean corpuscular volume (p = 0.0346) were found to be of prognostic importance. The 7 dogs in which white blood cell count of less than 2000 had shorter median survival time (3 days) than the remaining 28 dogs (8 days), but no statistical significance was found between leukopenic and survival. The distribution of packed cell volume and hemoglobin measurement was such that the overall risk of dying in the two groups was comparable. Further studies are needed to more accurately assess these results.

• 대한수의학회지
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• 제35권1호
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• pp.75-85
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• 1995

An indirect immunofluorescent antibody test was applied to survey the antibody prevalence on five canine viruses including canine distempervirus(CDV), canine parvovirus(CPV), canine coronavirus(CCV), canine adenovirus type-2(CAV-2), canine parainfluenzavirus(CPIV) in dogs. The period studied was from October 1992 to June 1993. A total of 80 dog sera was collected from veterinary clinics in Kwangju and Seoul, and examined for the presence of virus antibodies. Immunofluorescent antibodies(IFA) to all viruses were present in a high percentage of 80 sera tested. Seventyfive(93.8%) showed detectable IFA against CPV, 67(83.8%) against CDV, 51(63.8%) against CCV, 42(52.5%) against CPIV and 34(42.5%) against CAV-2. These suggested that all viruses were endemic in the communities. IFA levels against each virus were also distributed fairly irregularly. IFAs for CDV and CPV were detected more frequently with a relatively high incidence in vaccinated group less than 1 years of age. IFAs for CAV-2 were detected more frequently with growing age. In the correlation of clinical signs and antibody prevalence, dogs that showed hematochezia and vomiting had high titers in the positive sera is noteworthy, particularly for CDV and CPV. The significance between dogs those who had diarrhea, dyspnea and salivation and those viruses were obscure.

• Journal of Microbiology and Biotechnology
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• 제21권11호
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• pp.1109-1115
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• 2011

A highly sensitive and specific TaqMan real-time PCR was used to quantify hepatopancreatic parvovirus (HPV) type III infections in wild broodstocks and hatchery-reared postlarvae (PL) of Fenneropenaeus chinensis. Totals of 159 and 162 wild brooders from three locations were captured, and 140 and 180 PL were obtained from seven and six commercial hatcheries in 2007 and 2008, respectively. Among the three wild broodstock groups from 2007, only 1 group showed HPV infection and 3.2% of 159 brooders were positive for HPV infection. In 2008, HPV infections were observed from all three wild broodstock groups with $1.93{\times}10^4$ copies/mg tissue of pleopods. Of 162 brooders, 26.6% were positive for HPV infection. No PL from the two hatcheries collected in 2007 showed HPV infection, and PL from the rest of the five hatcheries had up to $1.74{\times}10^6$ copies/ng of DNA, and PL from three hatcheries showed HPV infections with over 1,000 copies/ng of DNA. The PL from all seven hatcheries collected in 2008 showed up to $2.10{\times}10^5$ HPV copies/ng of DNA. PL from two hatcheries showed less than 100 copies/ng of DNA, but PL from the rest of the hatcheries showed HPV infections with over 1,000 copies/ng of DNA. These results show that HPV type III is widely distributed in Korea in addition to previously reported HPV type I, and they can be effectively detected by type-specific realtime PCR.

• Journal of Veterinary Science
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• 제22권1호
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• pp.1.1-1.12
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• 2021

Background: Goslings in several Taiwanese farms experienced gosling feather loss disease (GFL) at 21-35 days and goose broke feather disease (GBF) at 42-60 days. The prevalence ranges from a few birds to 500 cases per field. It is estimated that about 12,000 geese have been infected, the morbidity is 70-80% and the mortality is 20-30%. Objectives: This study aims to investigate the pathogens that cause GFL and GBF. Focus on the study of the correlation between goose circovirus (GoCV) and goose parvovirus (GPV) with the goose feather loss in southern Taiwan. Furthermore, a phylogenetic tree was established to align the differences between southern and northern Taiwan and compare with virus strains from China and Europe. Methods: Samples were collected from animal hospitals. Molecular and microscopy diagnostics were used to examine 92 geese. Specific quantitative polymerase chain reaction (Q-PCR) assays are performed to evaluate GPV and GoCV viral loads and simultaneously evaluated the feather loss conditions in geese with the scoring method. Results: High prevalence of GoCV and GPV infection in geese showing signs of GFL and GBF. Inclusion body was detected in the feather follicles and Lieberkuhn crypt epithelial cells. The Q-PCR showed the high correlation between feather loss and viruses during 3rd-5th week. However, the infection was not detected using the same test in 60 healthy geese. Conclusions: Thus, GFL and GBF appear to be significantly closely related to GoCV and GPV. The geese feathers showed increasing recovery after being quarantined and disinfected.

• Journal of Microbiology and Biotechnology
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• 제29권9호
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• pp.1391-1400
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• 2019

Canine parvoviral enteritis (PVE) is an important intestinal disease of the puppies; however, the potential impact of the canine parvovirus (CPV) on the gut microbiota has not been investigated. Therefore, the aim of this study was to evaluate the gut microbial shifts in puppies naturally infected with CPV. Fecal samples were collected from healthy dogs and those diagnosed with PVE at 4, 6, 8, and 12 weeks of age. The distal gut microbiota of dogs was characterized using Illumina MiSeq sequencing of the bacterial 16S rRNA genes. The sequence data were analyzed using QIIME with an Operational Taxonomic Unit definition at a similarity cutoff of 97%. Our results showed that the CPV was associated with significant microbial dysbiosis of the intestinal microbiota. Alpha diversity and species richness and evenness in dogs with PVE decreased compared to those of healthy dogs. At the phylum level, the proportion of Proteobacteria was significantly enriched in dogs with PVE while Bacteroidetes was significantly more abundant in healthy dogs (p < 0.05). In dogs with PVE, Enterobacteriaceae was the most abundant bacterial family accounting for 36.44% of the total bacterial population compared to only 0.21% in healthy puppies. The two most abundant genera in healthy dogs were Prevotella and Lactobacillus and their abundance was significantly higher compared to that of dogs with PVE (p < 0.05). These observations suggest that disturbances of gut microbial communities were associated with PVE in young dogs. Evaluation of the roles of these bacterial groups in the pathophysiology of PVE warrants further studies.

• Journal of Veterinary Science
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• 제21권3호
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• pp.50.1-50.16
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• 2020

Background: Porcine parvovirus (PPV) is a single-stranded DNA virus that causes porcine reproductive failure. It is of critical importance to study PPV pathogenesis for the prevention and control of the disease. NS1, a PPV non-structural protein, is participated in viral DNA replication, transcriptional regulation, and cytotoxicity. Our previous research showed that PPV can activate nuclear factor kappa B (NF-κB) signaling pathway and then up-regulate the expression of interleukin (IL)-6. Objectives: Herein, the purpose of this study is to determine whether the non-structural protein NS1 of PPV also has the same function. Methods: Real-time quantitative reverse transcription polymerase chain reaction (RT-qPCR), enzyme-linked immunosorbent assay, western blot, immunofluorescence assay and small interfering RNA (siRNA) were used. Results: Our findings demonstrated that PPV NS1 protein can up-regulate the expression levels of IL-6 and tumor necrosis factor-alpha in a dose-dependent manner. Moreover, PPV NS1 protein was found to induce the phosphorylation of IκBα, then leading to the phosphorylation and nuclear translocation of NF-κB. In addition, the NS1 protein activated the upstream pathways of NF-κB. Meanwhile, TLR2-siRNA assay showed TLR2 plays an important role in the activation of NF-κB signaling pathway induced by PPV-NS1. Conclusions: These findings indicated that PPV NS1 protein induced the up-regulated of IL-6 expression through activating the TLR2 and NF-κB signaling pathways. In conclusion, these findings provide a new avenue to study the innate immune mechanism of PPV infection.

• Pediatric Infection and Vaccine
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• 제28권3호
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• pp.181-188
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• 2021

신장이식 환자에서 만성 빈혈은 치료 과정 중 발생하는 합병증으로 바이러스 감염, 적혈구형성호르몬결핍, 출혈, 면역억제제의 부작용 등과 관련이 있다. 신장이식 환자에서 파르보바이러스 B19에 의한 빈혈은 초감염 혹은 기존 감염의 재활성화로 발생할 수 있으며, 아직 이에 대한 치료법은 정립이 되어 있지 않다. 특히, 소아청소년 신장이식 환자에 대한 보고나 자료는 더욱 드물다. 저자들은 소아청소년 신장이식 환자에서 발생한 심각한 빈혈을 파르보바이러스 B19 PCR 검사법으로 진단하고, 정맥내글로불린으로 재발 없이 성공적으로 치료한 두 증례와 문헌 고찰을 보고하는 바이다.

• Journal of Veterinary Science
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• 제23권1호
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• pp.14.1-14.11
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• 2022

Background: Carnivore protoparvovirus 1, also known as canine parvovirus type 2 (CPV-2), is the main pathogen in hemorrhagic gastroenteritis in dogs, with a high mortality rate. Three subtypes (a, b, c) have been described based on VP2 residue 426, where 2a, 2b, and 2c have asparagine, aspartic acid, and glutamic acid, respectively. Objectives: This study examined the presence of CPV-2 variants in the fecal samples of dogs diagnosed with canine parvovirus in Bogotá. Methods: Fecal samples were collected from 54 puppies and young dogs (< 1 year) that tested positive for the CPV through rapid antigen test detection between 2014-2018. Molecular screening was developed for VP1 because primers 555 for VP2 do not amplify, it was necessary to design a primer set for VP2 amplification of 982 nt. All samples that were amplified were sequenced by Sanger. Phylogenetics and structural analysis was carried out, focusing on residue 426. Results: As a result 47 out of 54 samples tested positive for VP1 screening, and 34/47 samples tested positive for VP2 980 primers as subtype 2a (n = 30) or 2b (n = 4); subtype 2c was not detected. All VP2 sequences had the amino acid, T, at 440, and most Colombian sequences showed an S514A substitution, which in the structural modeling is located in an antigenic region, together with the 426 residue. Conclusions: The 2c variant was not detected, and these findings suggest that Colombian strains of CPV-2 might be under an antigenic drift.