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The study was performed to develop a non-culture method to quantify viable loads of  Lactobacillus acidophilus  NCFM using RNA-based molecular technique. The ‘growth curve’ and ‘cycle threshold curve’ were developed respectively by plate counting and using cycle threshold (C T ) values. ‘Standard curve’ was constructed using cells per milliliter and relative C T  values. A maximum viable count (1.5 ± 0.15) × 10 10  cells/mL with a minimum C T  value 20.18 ± 0.56 was achieved following 18 h of growth. The two parameters were inversely proportional to each other over the exponential growth. The ‘standard curve’ corresponded to equation y = 2E + 28e −2.034x  (y = cells/mL, x = C T  value; R 2  = 0.993), and no sample showed significant difference between ‘plate count’ and relative ‘PCR count’ following the validation process. Industrial adaptation of this method in dairy processing could potentially contribute to a faster enumeration of viable  L. acidophilus  NCFM compared to plate counting.

A Culture-Independent Method for Enumeration of Viable Load of Lactobacillus acidophilus NCFM by using Real-Time PCR