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Development of a Domestic Alternative Positive Control Strain to Bacillus cereus ATCC 14579 for Microbial Assays
Su-Hyeon Joung1, Yu-Si Lee1, Byeong Joon Kim1
1Division of Food Microbiology, National Institute of Food and Drug Safety Evaluation, Ministry of Food and Drug Safety, Cheongju, Republic of Korea.
Abstract:
Most positive control strains in microbial assays are sourced from abroad. Bacillus cereus ATCC 14579 is used as the positive control strain in microbial assays described in the Manual for the Detection of Foodborne Pathogens at Outbreaks and the FDA's Bacteriological Analytical Manual method. This study aimed to identify a domestically isolated strain from the Ministry of Food and Drug Safety (MFDS) to replace B. cereus ATCC 14579. A total of 323 strains from the MFDS were collected, and gene-targeted polymerase chain reaction and biochemical tests were performed to characterize candidate strains. Sixty-one candidate strains showing identical biochemical and molecular characteristics to ATCC 14579 were further subjected to whole-genome sequencing. Comparative genetic analyses using MLST, SNP distance, and OrthoANI revealed that B. cereus MFDS 1004972 shared the same sequence type, exhibited SNP distance of fewer than 10 SNPs, and showed 100% ANI with ATCC 14579. Therefore, B. cereus MFDS 1004972 is proposed as a suitable domestic alternative to the imported strain.
Insights
A domestic Bacillus cereus strain, MFDS 1004972, has been identified as a suitable alternative to the imported ATCC 14579 strain for microbial assays. This finding supports the use of local microbial control strains in foodborne pathogen detection.
Area of Science:
- Microbiology
- Food Safety
- Genomics
Background:
- Positive control strains for microbial assays are often imported.
- Bacillus cereus ATCC 14579 is a commonly used control strain in foodborne pathogen detection methods.
- There is a need for domestically sourced control strains.
Purpose of the Study:
- To identify a domestic Bacillus cereus strain to replace the imported ATCC 14579.
- To ensure the availability of reliable control strains for microbial assays within the country.
Main Methods:
- Screening of 323 domestic strains from the Ministry of Food and Drug Safety (MFDS).
- Utilized gene-targeted polymerase chain reaction (PCR) and biochemical tests for initial characterization.
- Whole-genome sequencing and comparative genomic analyses (MLST, SNP distance, OrthoANI) for detailed comparison.
Main Results:
- Identified 61 candidate strains with similar characteristics to ATCC 14579.
- Bacillus cereus MFDS 1004972 demonstrated identical sequence type, <10 SNP distance, and 100% Orthogonal Average Nucleotide Identity (OrthoANI) to ATCC 14579.
- MFDS 1004972 is genetically highly similar to the reference strain.
Conclusions:
- Bacillus cereus MFDS 1004972 is proposed as a suitable domestic alternative to Bacillus cereus ATCC 14579.
- This domestic strain can support national food safety and microbial testing capabilities.
- Facilitates greater self-sufficiency in microbial assay controls.
