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) 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.

Airway obstruction during general anesthesia in a premature infant suspecting bronchospasm and/or airway malacia: A case report

Airway management is challenging during general anesthesia particularly in small infants. Airway obstruction is prone to occur in premature infants during general anesthesia due to several reasons. We report a case of airway obstruction occurred during the induction of general anesthesia in a 2-month-old infant. Several attempts at endotracheal intubation with positive pressure ventilation resulted in repeated patterns of no end-tidal carbon dioxide output after each trial of endotracheal intubation, but it was reappeared after extubation. However, anesthetic induction with self-respiration and gentle assistance with manual bagging led to a successful intubation. This case was explained by hydromechanics in a collapsible premature airway.