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Detection and absolute quantification of Lactiplantibacillus plantarum ATCC 202195 by quantitative real-time PCR
Jakaria Shawon1, Lisa G Pell2, Mamun Kabir3
1Nutrition and Clinical Services Division, International Centre for Diarrhoeal Disease Research, Bangladesh (icddr,b) , Dhaka, Bangladesh.
Insights
A new real-time PCR assay can detect and quantify the probiotic Lactiplantibacillus plantarum ATCC 202195. This method aids in monitoring probiotic safety and ensuring intervention quality for infant health.
Area of Science:
- Microbiology
- Molecular Biology
- Probiotics
Background:
- Probiotic interventions, including Lactiplantibacillus plantarum ATCC 202195 plus fructooligosaccharide (FOS), show promise in reducing infant infections like sepsis and respiratory illnesses.
- Strain-specific detection and quantification of probiotics are crucial for monitoring safety (e.g., bacteremia) and ensuring the quality of probiotic products.
Purpose of the Study:
- To develop and optimize a specific and sensitive real-time PCR assay for the detection and quantification of Lactiplantibacillus plantarum ATCC 202195.
- To provide a molecular tool for assessing the safety and quality of probiotic interventions using this specific strain.
Main Methods:
- Design of specific primer and probe sequences tailored for Lactiplantibacillus plantarum ATCC 202195.
- Development and optimization of a real-time quantitative PCR (qPCR) assay.
Main Results:
- Successfully designed and validated primer and probe sequences for high-specificity detection.
- Optimized a real-time PCR assay capable of accurately detecting and quantifying Lactiplantibacillus plantarum ATCC 202195.
- Achieved high sensitivity in the developed assay.
Conclusions:
- A novel, highly specific, and sensitive real-time PCR assay for Lactiplantibacillus plantarum ATCC 202195 has been established.
- This assay provides a critical tool for quality control and safety monitoring of probiotic interventions involving this strain.
- Facilitates further clinical research and application of L. plantarum ATCC 202195 in infant health.
Importance:
When administered for seven consecutive days shortly after birth, the probiotic bacterium Lactiplantibacillus plantarum ATCC 202195 plus fructooligosaccharide (FOS) was reported to reduce sepsis and lower respiratory tract infection events during early infancy in a randomized trial in India. Since probiotic effects are often strain specific, strain-level detection and quantification by routine molecular methods enables the monitoring of safety outcomes, such as probiotic-associated bacteremia, and allows for the quality of probiotic interventions to be assessed and monitored (i.e., verify strain identity and enumerate). Despite the potential clinical applications of L. plantarum ATCC 202195, an assay to detect and quantify this strain has not previously been described. Herein, we report the design of primer and probe sequences to detect L. plantarum ATCC 202195 and the development and optimization of a real-time PCR assay to detect and quantify the strain with high specificity and high sensitivity.
