Established qPCR Method Using Strain-Specific Primers for Enumeration of Paraprobiotic Lactobacillus plantarum L137

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Published: Sep 26, 2024
DOI: https://doi.org/10.47191/ijpbms/v4-i9-09
Keywords:
Lactobacillus plantarum, paraprobiotic, real-time PCR, quantification, validation

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Yen Thi Ngoc Nguyen
Faculty of Pharmacy, Nguyen Tat Thanh University, 298-300 Nguyen Tat Thanh Street, Ward 13, District 4, Ho Chi Minh City, 700000, Vietnam
Nhan Thi Vo
Faculty of Pharmacy, Nguyen Tat Thanh University, 298-300 Nguyen Tat Thanh Street, Ward 13, District 4, Ho Chi Minh City, 700000, Vietnam

Abstract

Lactobacillus plantarum, extensively present in fermented foods and the human intestinal tract, has numerous health advantages, including regulating systemic immunity, maintaining a balanced intestinal flora, and lowering the risk of tumour progression. Because the health benefits and safety are strain-specific, it is essential to have a trustworthy method for the specific detection and enumeration of L. plantarum L137 to verify probiotic efficacy. The real-time PCR tool was developed using strain-specific primer pairs from the specialized plasmid pLTK2 and successfully validated for specificity, precision, and amplification efficiency. The assay achieved 100% true positive and 0% false positive results by effectively amplifying the target sample and not amplifying any of the six non-target strains. The limit of quantification was established to be 1.7.103 CFU/reaction. The reaction efficiency values were 86.7%, with an R2 value of 0.9864. The repeatability's relative standard deviation (RSD) was 2.34%. The qPCR assay for L. plantarum L137 was found to meet the specific, precise, and efficient requirements. To meet compliance standards, it is crucial to have a reliable and approved assay for strain-specific identification and enumeration for probiotic products.

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How to Cite
Yen Thi Ngoc Nguyen, & Nhan Thi Vo. (2024). Established qPCR Method Using Strain-Specific Primers for Enumeration of Paraprobiotic Lactobacillus plantarum L137. International Journal of Pharmaceutical and Bio Medical Science, 4(9), 759–764. https://doi.org/10.47191/ijpbms/v4-i9-09
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Vol. 4 No. 9 (2024): Volume 04 Issue 09 September 2024
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This work is licensed under a Creative Commons Attribution 4.0 International License.

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