• Korean Journal of Microbiology
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• v.39 no.1
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• pp.62-65
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• 2003

A procedure to increase the stability of 6B capsular polysaccharide on microbead surface in the mutibead assay, a serotyping method for Streptococcus pneumoniae, was studied. Pneumococcal capsular polysaccharide 6B was conjugated to bvine serum albumin (BSA), and the coating efficiency and the stability of the 6B-BSA complex was measured. The 6B-BSA complex showed about 200-fold higher coating efficiency to polystyrene surface than 6B polysaccharide. And the stability of the 6B- BSA to be used in the multibead assay for 30 days after coating.

• Pediatric Infection and Vaccine
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• v.22 no.2
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• pp.97-105
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• 2015

Purpose: Serotyping pneumococcal isolates is important to monitor efficacy of pneumococcal vaccines. Because of difficulties of typing pnueumocci, a multiplex bead-based (multibead) serotyping assay was recently introduced. The aim of this study is to establish a new multibead serotyping assay and to apply this method to analyze clinical isolates of pneumococci in Korea. Methods: To establish the multibead serotyping assay, six key reagents were transferred from University of Alabama at Birmingham (UAB) to Ewha Center for Vaccine Evaluation and Study (ECVES): bead set coated with polysaccharide and monoclonal antibody pool were used in one multiplex inhibition-type immunoassay and 2 bead sets coated DNA probe and 2 primer pools were used in two multiplex PCR-based assays. After multibead serotyping assay was set up, 75 test samples of pneumococci were analyzed whether ECVES is able to identify serotype correctly. After confirming the performance, serotyping assay was applied to identify serotypes of 528 clinical isolates of pneumococci collected from 3 different hospitals. Results: After establishment of the multibead pneumococcal serotyping assay system at ECVES, 75 test samples were analyzed. There was no discrepancy of serotypes of 75 test samples between the results assigned at UAB and those at ECVES. The serotypes of 528 pneumococci isolated from patients or healthy subjects were determined in 94.3% of isolates (498/528). Conclusions: The multibead pneumococcal serotyping assay can be successfully established in Korea. With this method, surveillance of serotypes of pneumococci isolated from patients as well as healthy subjects could be studied.

• Clinical and Experimental Pediatrics
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• v.50 no.2
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• pp.151-156
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• 2007

Purpose : Streptococcus pneumoniae is a major etiologic agent for pneumonia, meningitis, otitis media, and sepsis among young children. Multi-drug resistant strains have raised great concern worldwide, thus the importance of prevention with vaccines has been emphasized. However, vaccines may force the appearance of pneumococcal infections by nonvaccine serotypes. Thus, distribution of pneumococcal serotypes should be monitored to estimate vaccine efficacy. We used a new and efficient multibead assay in determining pnemococcal serotypes. Methods : From January to February 2005, 643 children were recruited from ten day care centers to isolate pneumococci from their oropharynx. Pneumococcal serotyping was performed on 62 pneumococcal isolates from 60 children by multibead assay. This immunoassay required two sets of latex particles coated with pneumococcal polysaccharides and serotype-specific antibodies. Twenty four newly developed monoclonal antibodies specific for common serotypes and a pool of polyclonal rabbit sera for some of the less common serotypes were used. Results : The most prevalent pneumococcal serotypes were serotype 6A, 19A, 19F, 23F, and 11A/D/F which accounted more than 50 precent of all the 62 pneumococcal isolates. We found that multibead assay can be performed very rapidly and objectively. Conclusion : This multibead immunoassay was very useful in serotyping clinical isolates of S. pneumoniae because it was simple, reliable and fast.