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Development of a New Approach to Determine the Potency of Bacille Calmette–Guérin Vaccines Using Flow Cytometry
OBJECTIVES: To circumvent the limitations of the current golden standard method, colony-forming unit (CFU) assay, for viability of Bacille Calmette–Guérin (BCG) vaccines, we developed a new method to rapidly and accurately determine the potency of BCG vaccines. METHODS: Based on flow cytometry (FACS...
Autores principales: | , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
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Korea Centers for Disease Control and Prevention
2017
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Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5749480/ https://www.ncbi.nlm.nih.gov/pubmed/29354397 http://dx.doi.org/10.24171/j.phrp.2017.8.6.06 |
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author | Gweon, Eunjeong Choi, Chanwoong Kim, Jaeok Kim, Byungkuk Kang, Hyunkyung Park, Taejun Ban, Sangja Bae, Minseok Park, Sangjin Jeong, Jayoung |
author_facet | Gweon, Eunjeong Choi, Chanwoong Kim, Jaeok Kim, Byungkuk Kang, Hyunkyung Park, Taejun Ban, Sangja Bae, Minseok Park, Sangjin Jeong, Jayoung |
author_sort | Gweon, Eunjeong |
collection | PubMed |
description | OBJECTIVES: To circumvent the limitations of the current golden standard method, colony-forming unit (CFU) assay, for viability of Bacille Calmette–Guérin (BCG) vaccines, we developed a new method to rapidly and accurately determine the potency of BCG vaccines. METHODS: Based on flow cytometry (FACS) and fluorescein diacetate (FDA) as the most appropriate fluorescent staining reagent, 17 lots of BCG vaccines for percutaneous administration and 5 lots of BCG vaccines for intradermal administration were analyzed in this study. The percentage of viable cells measured by flow cytometry along with the total number of organisms in BCG vaccines, as determined on a cell counter, was used to quantify the number of viable cells. RESULTS: Pearson correlation coefficients of FACS and CFU assays for percutaneous and intradermal BCG vaccines were 0.6962 and 0.7428, respectively, indicating a high correlation. The coefficient of variation value of the FACS assay was less than 7%, which was 11 times lower than that of the CFU assay. CONCLUSION: This study contributes to the evaluation of new potency test method for FACS-based determination of viable cells in BCG vaccines. Accordingly, quality control of BCG vaccines can be significantly improved. |
format | Online Article Text |
id | pubmed-5749480 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2017 |
publisher | Korea Centers for Disease Control and Prevention |
record_format | MEDLINE/PubMed |
spelling | pubmed-57494802018-01-19 Development of a New Approach to Determine the Potency of Bacille Calmette–Guérin Vaccines Using Flow Cytometry Gweon, Eunjeong Choi, Chanwoong Kim, Jaeok Kim, Byungkuk Kang, Hyunkyung Park, Taejun Ban, Sangja Bae, Minseok Park, Sangjin Jeong, Jayoung Osong Public Health Res Perspect Original Article OBJECTIVES: To circumvent the limitations of the current golden standard method, colony-forming unit (CFU) assay, for viability of Bacille Calmette–Guérin (BCG) vaccines, we developed a new method to rapidly and accurately determine the potency of BCG vaccines. METHODS: Based on flow cytometry (FACS) and fluorescein diacetate (FDA) as the most appropriate fluorescent staining reagent, 17 lots of BCG vaccines for percutaneous administration and 5 lots of BCG vaccines for intradermal administration were analyzed in this study. The percentage of viable cells measured by flow cytometry along with the total number of organisms in BCG vaccines, as determined on a cell counter, was used to quantify the number of viable cells. RESULTS: Pearson correlation coefficients of FACS and CFU assays for percutaneous and intradermal BCG vaccines were 0.6962 and 0.7428, respectively, indicating a high correlation. The coefficient of variation value of the FACS assay was less than 7%, which was 11 times lower than that of the CFU assay. CONCLUSION: This study contributes to the evaluation of new potency test method for FACS-based determination of viable cells in BCG vaccines. Accordingly, quality control of BCG vaccines can be significantly improved. Korea Centers for Disease Control and Prevention 2017-12 2017-12-31 /pmc/articles/PMC5749480/ /pubmed/29354397 http://dx.doi.org/10.24171/j.phrp.2017.8.6.06 Text en Copyright ©2017, Korea Centers for Disease Control and Prevention http://creativecommons.org/licenses/by-nc-nd/4.0/ This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/). |
spellingShingle | Original Article Gweon, Eunjeong Choi, Chanwoong Kim, Jaeok Kim, Byungkuk Kang, Hyunkyung Park, Taejun Ban, Sangja Bae, Minseok Park, Sangjin Jeong, Jayoung Development of a New Approach to Determine the Potency of Bacille Calmette–Guérin Vaccines Using Flow Cytometry |
title | Development of a New Approach to Determine the Potency of Bacille Calmette–Guérin Vaccines Using Flow Cytometry |
title_full | Development of a New Approach to Determine the Potency of Bacille Calmette–Guérin Vaccines Using Flow Cytometry |
title_fullStr | Development of a New Approach to Determine the Potency of Bacille Calmette–Guérin Vaccines Using Flow Cytometry |
title_full_unstemmed | Development of a New Approach to Determine the Potency of Bacille Calmette–Guérin Vaccines Using Flow Cytometry |
title_short | Development of a New Approach to Determine the Potency of Bacille Calmette–Guérin Vaccines Using Flow Cytometry |
title_sort | development of a new approach to determine the potency of bacille calmette–guérin vaccines using flow cytometry |
topic | Original Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5749480/ https://www.ncbi.nlm.nih.gov/pubmed/29354397 http://dx.doi.org/10.24171/j.phrp.2017.8.6.06 |
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