期刊
JOURNAL OF VIROLOGICAL METHODS
卷 208, 期 -, 页码 66-78出版社
ELSEVIER SCIENCE BV
DOI: 10.1016/j.jviromet.2014.07.030
关键词
Arbovirus; Antigen; Immunoglobulin M antibody-capture enzyme-linked immunosorbent assay (MAC-ELISA); Beta-propiolactone; Gamma-irradiation
资金
- Intramural CDC HHS [CC999999] Funding Source: Medline
Arboviruses are medically important pathogens that cause human disease ranging from a mild fever to encephalitis. Laboratory diagnosis is essential to differentiate arbovirus infections from other pathogens with similar clinical manifestations. The Arboviral Diseases Branch (ADB) reference laboratory at the CDC Division of Vector-Borne Diseases (DVBD) produces reference antigens used in serological assays such as the virus-specific immunoglobulin M antibody-capture enzyme-linked immunosorbent assay (MAC-ELISA). Antigen production in cell culture has largely replaced the use of suckling mice; however, the methods are not directly transferable. The development of a cell culture antigen production algorithm for nine arboviruses from the three main arbovirus families, Flaviviridae, Togaviridae, and Bunyaviridae, is described here. Virus cell culture growth and harvest conditions were optimized, inactivation methods were evaluated, and concentration procedures were compared for each virus. Antigen performance was evaluated by the MAC-ELISA at each step of the procedure. The antigen production algorithm is a framework for standardization of methodology and quality control; however, a single antigen production protocol was not applicable to all arboviruses and needed to be optimized for each virus. Published by Elsevier B.V.
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