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Direct On-Chip Diagnostics of Streptococcus bovis/Streptococcus equinus Complex in Bovine Mastitis Using
Jaewook Kim1, Eiseul Kim1, Seung-Min Yang1
1Institute of Life Sciences & Resources, Department of Food Science and Biotechnology, Kyung Hee University, Yongin 17104, Republic of Korea.
Insights
A new portable diagnostic tool rapidly detects the Streptococcus bovis/Streptococcus equinus complex (SBSEC) in livestock and food. This method offers quick, on-site detection, enhancing animal health and food safety.
Area of Science:
- Veterinary Microbiology
- Genomics
- Biosensor Technology
Background:
- The Streptococcus bovis/Streptococcus equinus complex (SBSEC) poses risks to livestock health and food safety.
- Accurate identification of SBSEC species is challenging due to misclassifications in genomic data.
- Existing diagnostic methods can be time-consuming and require laboratory settings.
Purpose of the Study:
- To develop and validate an innovative on-site diagnostic method for rapid SBSEC detection.
- To identify genetic markers for improved SBSEC species classification.
- To provide a portable and efficient alternative to traditional laboratory diagnostics.
Main Methods:
- Comprehensive genomic analysis of 206 SBSEC genomes to identify discriminatory genetic markers.
- Development of a portable quantitative polymerase chain reaction (qPCR) assay incorporating identified markers.
- Field validation of the portable qPCR system using samples from cattle udders, farm environments, and dairy products.
Main Results:
- Identified genetic markers that enhance SBSEC classification accuracy, resolving misclassifications.
- Developed a portable qPCR system capable of detecting SBSEC species with high sensitivity (down to 10^1 CFU/mL).
- Achieved rapid on-site diagnosis within 30 minutes, with performance comparable to laboratory methods.
- Validated the method in real-world conditions, detecting mastitis-associated bacteria in 51 out of 100 samples.
Conclusions:
- The developed portable qPCR offers a rapid, sensitive, and reliable on-site diagnostic solution for SBSEC.
- This technology is suitable for immediate use in clinical, agricultural, and environmental settings.
- It presents a practical alternative to whole-genome sequencing for early detection and management of SBSEC-related issues.
Abstract:
This study introduces an innovative on-site diagnostic method for rapidly detecting the Streptococcus bovis/Streptococcus equinus complex (SBSEC), crucial for livestock health and food safety. Through a comprehensive genomic analysis of 206 genomes, this study identified genetic markers that improved classification and addressed misclassifications, particularly in genomes labeled S. equinus and S. lutetiensis. These markers were integrated into a portable quantitative polymerase chain reaction (qPCR) that can detect SBSEC species with high sensitivity (down to 101 or 100 colony-forming units/mL). The portable system featuring a flat chip and compact equipment allows immediate diagnosis within 30 min. The diagnostic method was validated in field conditions directly from cattle udders, farm environments, and dairy products. Among the 100 samples, 51 tested positive for bacteria associated with mastitis. The performance of this portable qPCR was comparable to laboratory methods, offering a reliable alternative to whole-genome sequencing for early detection in clinical, agricultural, and environmental settings.
