A Culture-Independent Method for Enumeration of Viable Load of Lactobacillus acidophilus NCFM by using Real-Time PCR
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Md. Aslam Ali
Abstract
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 (CT) values. ‘Standard curve’ was constructed using cells per milliliter and relative CT values. A maximum viable count (1.5 ± 0.15) × 1010 cells/mL with a minimum CT 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 = CT value; R2 = 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.
Acknowledgments
The authors acknowledge the financial support of “National High-Level Talents Special Support Program of China” and “Academic Backbone” Project of Northeast Agricultural University (15XG26), Harbin, Heilongjiang, China for completion of this research project.
Conflicts of interest: The authors declare that they have no conflict of interest.
References
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Articles in the same Issue
- Critical Review
- Significance of Temperature and Humidity Control for Agricultural Products Storage: Overview of Conventional and Advanced Options
- Articles
- Safe and Complete Extraction of Astaxanthin from Haematococcus pluvialis by Efficient Mechanical Disruption of Cyst Cell Wall
- Analysis of Volatile Compounds from Wheat Flour in the Heating Process
- Alternative Tempering of Sugar-Free Dark Chocolates by βv Seeding: Sensorial, Micro-Structural and Some Physical Properties and Volatile Profile
- Numerical Modeling of Air Flow in a Cabinet Dryer Equipped by Deflector Plates
- Effects of Microwave and Cold Plasma Assisted Hydrodistillation on Lemon Peel Oil Extraction
- A Culture-Independent Method for Enumeration of Viable Load of Lactobacillus acidophilus NCFM by using Real-Time PCR
