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ARS Home » Southeast Area » Stoneville, Mississippi » Warmwater Aquaculture Research Unit » Research » Publications at this Location » Publication #416282
Research Project: Umbrella Project for Food Safety

Location: Warmwater Aquaculture Research Unit

Title: Development of a qPCR Detection Approach for Pathogenic Burkholderia cenocepacia Associated with Fresh Vegetables

Author
item LIU, AIXIN - Mississippi State University
item PHILLIPS, KATE - Mississippi State University
item JIA, JIAKUAN - Mississippi State University
item DENG, PENG - Mississippi State University
item Zhang, Dunhua
item CHANG, SAM - Mississippi State University
item LU, SHI-EN - Mississippi State University

Submitted to: Food Microbiology
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 7/3/2023
Publication Date: 7/14/2023
Citation: Liu, A., Phillips, K., Jia, J., Deng, P., Zhang, D., Chang, S., Lu, S. 2023. Development of a qPCR Detection Approach for Pathogenic Burkholderia cenocepacia Associated with Fresh Vegetables. Food Microbiology. 115:104333. https://doi.org/10.1016/j.fm.2023.104333.
DOI: https://doi.org/10.1016/j.fm.2023.104333

Interpretive Summary: The natural environment can harbor Burkholderia cepacia complex organisms, including the pathogenic B. cenocepacia. A reliable method for detecting B. cenocepacia in fresh food and the environment is currently lacking. This study developed a quantitative real-time PCR (qPCR) method for B. cenocepacia detection, validated on fresh vegetable samples. This qPCR method offers high sensitivity and specificity, promising for B. cenocepacia detection and epidemiological research on Burkholderia cepacia complex organisms in fresh vegetables.

Technical Abstract: Natural environment serves as a reservoir for Burkholderia cepacia complex organisms, including the highly transmissible opportunistic human pathogen B. cenocepacia. Currently, there is a lack of an effective and quantitative method for B. cenocepacia detection in fresh food and other environmental niches. A quantitative real-time PCR (qPCR) detection method for B. cenocepacia bacteria was established in this study and validated using artificially inoculated fresh vegetable samples. Genome-wide comparative methods were applied to identify target regions for the design of species-specific primers. Assay specificity was measured with 12 strains of closely related Burkholderia bacteria and demonstrated the primer pair BCF6/R6 were 100% specific for detection of B. cenocepacia. The described qPCR assay evaluated B. cenocepacia with a 2 pg µl-1 limit of detection and appropriate linearity (R2 = 0.999). In 50 samples of experimentally infected produce (lettuce, onion, and celery), the assay could detect B. cenocepacia as low as 2.6 × 102 cells in each sample equal to 1 g. The established qPCR method quantitatively detects B. cenocepacia with high sensitivity and specificity, making it a promising technique for B. cenocepacia detection and epidemiological research on B. cepacia complex organisms from fresh vegetables.