Cabezas Sheila, Rojas Gertrudis, Pavon Alequis, Bernardo Lidice, Castellanos Yinet, Alvarez Mayling, Pupo Maritza, Guillen Gerardo, Guzman Maria G
Department of Virology, PAHO/WHO Collaborating Center for the Study of Dengue and its Vector, "Pedro Kourí" Tropical Medicine Institute, Marianao, Havana, Cuba.
Arch Virol. 2009;154(7):1035-45. doi: 10.1007/s00705-009-0401-1. Epub 2009 Jun 6.
The current study shows the usefulness of dengue-3- and dengue-4-specific phage-displayed antibody fragments as tools for viral detection and serotyping in sera from infected individuals. C6/36 HT cells were inoculated with acute-phase sera from patients, and supernatants were collected daily and analyzed by ELISA using phage-displayed antibody fragments as serotype-specific detector reagents. Serotyping of most samples was possible as early as two to three days postinoculation. Results were comparable with those obtained by indirect immunofluorescence assay but were obtained in a shorter period of time (<1 week). Phage-displayed antibody fragments were better tools for diagnosis and serotyping than their soluble counterparts. Our approach combines the advantages of viral isolation and ELISA techniques. These results could be the basis for the development of a high-throughput method for identifying dengue virus serotypes, which is crucial for the management and control of the disease.
当前研究表明,登革3型和登革4型特异性噬菌体展示抗体片段可作为检测受感染个体血清中病毒及进行血清分型的工具。将患者急性期血清接种于C6/36 HT细胞,每天收集上清液,并使用噬菌体展示抗体片段作为血清型特异性检测试剂通过ELISA进行分析。大多数样本在接种后两到三天即可进行血清分型。结果与间接免疫荧光法相当,但所需时间更短(<1周)。噬菌体展示抗体片段比其可溶性对应物更适合用于诊断和血清分型。我们的方法结合了病毒分离和ELISA技术的优点。这些结果可为开发用于鉴定登革病毒血清型的高通量方法奠定基础,这对于该疾病的管理和控制至关重要。