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Flow Cytometry: Evolution of Microbiological Methods for Probiotics Enumeration.

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This study validated the ISO 19344:2015 flow cytometry method for quantifying Lactobacillus rhamnosus GG (ATCC 53103) in finished products. The analytical method proved reliable and robust for enumerating this probiotic.

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Area of Science:

  • Microbiology and Analytical Chemistry
  • Probiotic quantification and method validation

Background:

  • Flow cytometry offers a rapid, alternative method for microbial enumeration, distinguishing viable, damaged, and dead cells.
  • The International Standard ISO 19344-IDF 232:2015 provides a universal framework for quantifying lactic acid bacteria using flow cytometry.

Framework:

  • Validation of the analytical method ISO 19344:2015 (E)-IDF 232:2015 (E) for quantifying Lactobacillus rhamnosus GG (ATCC 53103).
  • Adherence to United States Pharmacopeia (USP) and International Council for Harmonisation (ICH) guidelines for method validation.

Implementation:

  • Analytical method validation was performed on three industrial batches of finished product containing L. rhamnosus GG.
  • Key validation parameters included accuracy, precision (repeatability), intermediate precision (ruggedness), specificity, limit of quantification, linearity, range, and robustness.

Implications:

  • The cytofluorimetric analysis demonstrated significant validity and robustness for enumerating L. rhamnosus GG in finished product formulations.
  • The validated ISO 19344:2015 (E)-IDF 232:2015 (E) standard is suitable for routine quantification of this specific probiotic.