• 제목/요약/키워드: Viral titer

검색결과 93건 처리시간 0.024초

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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    • 제11권3호
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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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국내 사육 꿩에서 분리된 뉴켓슬병 바이러스의 hemagglutinin-neuraminidase(HN) 유전자의 클론닝과 염기서열 분석 (Molecular cloning and nucleotide sequence of the gene encoding hemagglutinin-neuraminidase(HN) of Newcastle disease virus isolated from a diseased pheasant in Korea)

  • 장경수;곽길한;장승익;김지영;김태용;송영환;송희종;전무형
    • 한국동물위생학회지
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    • 제25권3호
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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.

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

  • 김민재;김소현;김성언;장미진;이현승;김영훈;한지환;김진택;장필상
    • Pediatric Infection and Vaccine
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    • 제22권2호
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    • pp.63-68
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    • 2015
  • 목적: 저자들은 2013년 경기 북부의 한 병원에서 경험한 소규모 홍역 유행의 양상을 알아보기 위해 본 연구를 시행하였다. 방법: 2013년 8월부터 10월까지 가톨릭대학교 의정부성모병원에서 홍역으로 확진된 환아 15명의 의무기록을 후향적으로 분석하였다. 결과: 홍역으로 진단된 환아는 모두 15명으로, 신생아 1명, 영아 11명, 유아 3명이었으며 역전사 중합 효소연쇄반응법(RT-PCR)과 혈청 내 홍역 IgM 검사로 확진하였다. 모든 환아는 Measles-Mumps-Rubella (MMR) 접종을 받지 않았다. 홍역으로 확진된 환아의 바이러스 유전형은 모두 B3이었다. 9명(60%)의 환아가 원내 감염에 의해 홍역에 걸렸다. 잠복기는 8-15일이었고 발열은 접촉 후 평균 10일째 발생하여 평균 8일 동안 39도 이상으로 지속되는 양상을 보였다. 발진은 접촉 후 평균 13일째 발생하였다. 40%의 환아가 호흡기 합병증을 보였고 53%의 환아가 설사를 동반하였다. 결론: 우리나라는 꾸준한 접종 사업을 통해 홍역 퇴치 수준에 이르게 되었지만 홍역 재유행, 특히 영아에 대한 위험성을 간과할 수 없게 되었다. 영유아에서 홍역이 의심될 때에는 가속접종, 면역글로불린 투여와 같은 처치를 적극적으로 시행하는 것이 필요하다. 향후 홍역 유행을 막기 위한 첫 번째 단계로, 신생아, 영아, 가임기 여성의 홍역 IgG 항체가 재조사가 필요할 것으로 사료된다.