• Title/Summary/Keyword: viral titer

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Removal and Inactivation of Viruses during Manufacture of a High Purity Antihemophilic Factor VII Concentration from Human Plasma

  • Kim, In-Seop;Choi, Yong-Woon;Lee, Sung-Rae;Woo, Hang-Sang;Lee, Soung-Min
    • Journal of Microbiology and Biotechnology
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    • v.11 no.3
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    • pp.497-503
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    • 2001
  • The purpose of this study was to examine the efficacy and mechanism of the cryo-precipitation, solvent/detergent (S/D) treatment, monoclonal anti-FVIIIc antibody (mAb) column chromatography, Q-Sepharose column chromatography, and lyophilization involved in the manufacture of antithemophilic factor VII(GreenMono) from human plasma, in the removal and/or inactivation of blood-borne viruses. A variety of experimental model viruses for human pathogenic viruses, including the bovine viral diarrhoea virus (BVDV), bovine herpes virus (BHV), murine encephalomyocarditis virus (EMCV), and porcine parvovirus (PPV), were all selected for this study. BHV and EMCV were effectively partitioned from a factor VII during the cryo-precipitation with a log reduction factor of 2.83 and 3.24, respectively. S/D treatment using the organic solvent, tri(n-butyl) phosphate (TNBP), and the detergent, Triton X-100, was a robust and effective step in inactivating enveloped viruses. The titers of BHV and BVDV were reduced from the initial titer of 8.85 and $7.89{log_10} {TCID_50}$, respectively, reaching undetectable levels within 1 min of the S/D treatment. The mAb chromatography was the most effective step for removing nonenveloped viruses, EMCV and PPV, with the log reduction factors of 4.86 and 3.72, respectively. Q-Sepharose chromatography showed a significant efficacy for partitioning BHV, BVDV, EMCV, and PPV with the log reduction the log reduction factors of 2.32, 2.49, 2.60, and 1.33 respectively. Lyophilization was an effective step in inactivating g nonenveloped viruses rather than enveloped viruses, where the log reduction factors of BHV, BVDV, DMCV, and PPV were 1.41, 1.79, 4.76, and 2.05, respectively. The cumulative log reduction factors of BHV, BVDV, EMCV, and PPV were ${\geqq}$11.12, ${\geqq}$7.88, 15.46, and 7.10, respectively. These results indicate that the production process for GreenMono has a sufficient virus-reducing capacity to achieve a high margin of the virus safety.

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Molecular cloning and nucleotide sequence of the gene encoding hemagglutinin-neuraminidase(HN) of Newcastle disease virus isolated from a diseased pheasant in Korea (국내 사육 꿩에서 분리된 뉴켓슬병 바이러스의 hemagglutinin-neuraminidase(HN) 유전자의 클론닝과 염기서열 분석)

  • 장경수;곽길한;장승익;김지영;김태용;송영환;송희종;전무형
    • Korean Journal of Veterinary Service
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    • v.25 no.3
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    • pp.245-257
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    • 2002
  • The gene encoding the HN protein from the CBP-1 strain, a heat stable Newcastle disease virus (NDV) isolated from diseased pheasants in Korea, was characterized by reverse transcriptase- polymerase chain reaction(RT-PCR) and the nucleotide and amino acid sequences were analyzed following cloning of the HN gene. In all of the NDV strains studied, a 1.75 kb size cDNA fragment for the HN gene was generated by RT-PCR and smaller specific band sizes harboring the internal portions of the HN gene were also detected by using four pairs of primers. The RT-PCR was sensitive enough to detect viral transcripts when the virus titer was above 25 hemagglutination units. The amplified 1.75 kb cDNA was cloned into a BamHI site of the pVL1393 Baculo transfer vector. The nucleotide sequences of the 1,758 bp HN gene from the CBP-1 strain were determined by the dye terminator cyclic sequencing method. The gene sequences were compared among the strains of CBP-1, Texas GB, Beaudette C, LaSota, B1 and Ulster. The homology of the CBP-1 HN gene to other HN variants was 97.8% to Texas GB, 98.4% to Beaudette C, 95.4% to LaSota, 95.6% to B1 and 90.2% to Ulster. As the deduced 577 amino acid sequences were compared among the strains, the homology for CBP-1 HN appeared to be 96.7% to Texas GB, 97.9% to Beaudette C, 95.5% to LaSota, 95.5% to B1 and 92.7% to Ulster. It was evident that the amino acid sequences included 5 sites for N-asparagine linked glycosylation and 12 cysteine residues. The three conserved leucine residues within the predicted transmembrane domain of the HN protein are amino acid 30, 37 and 44. The three antigenic sites on the HN protein of NDV are amino acids 347(Glu), 481(Asn) and 495(Glu). These data indicate that the genotype of the CBP-1 strain is more closely associated with the strains of Texas GB and Beaudette C than it is for the LaSota, B1 and Ulster strains.

A Small Outbreak of Measles in 2013: In a Single Hospital in Northern Gyeonggi-do (2013년 경기 북부 한 병원에서 확인된 소규모 홍역 유행)

  • Kim, Min Jae;Kim, So Hyun;Kim, Sung Un;Jang, Mi Jin;Lee, Hyun Seung;Kim, Young Hoon;Han, Ji Whan;Kim, Jin Tack;Jang, Pil Sang
    • Pediatric Infection and Vaccine
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    • v.22 no.2
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    • pp.63-68
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    • 2015
  • Purpose: This study analyzed a small outbreak of measles at a single hospital located in northern Gyeonggi-do in 2013. Methods: We reviewed the medical records of measles patients at The Catholic University of Korea Uijeongbu St. Mary's Hospital from August to October, 2013. Results: Fifteen children were confirmed to have measles by RT-PCR and serum IgM test; 1 neonate, 11 infants, and 3 toddlers. None of the patients had received Measles-Mumps-Rubella vaccination. All patients showed B3 type in viral genotyping. Nine children (60%) had been exposed to measles during treatment for other diseases in the pediatric ward. Incubation period was between 8 and 15 days. Fever started at a median 10 days after exposure and persisted for a median of 8 days. Rash showed at a median 13 days after exposure. Respiratory complications were observed in 40% of patients. Diarrhea developed in 53% of patients. Conclusion: Although measles has been well-controlled due to the high rate of vaccination coverage, it is possible to have an outbreak at any given time, especially in infants. We must learn from this outbreak, and remain fully aware of the possibility of reemergence and provide proper management, including vaccination or immune globulin administration, to infants exposed to measles. Reevaluation of serum IgG titer of neonates, infants, and pregnant women may be the first step to prevent further outbreaks.