Viable bacteria in the blood, i.e., bacteremia, can lead to bloodstream infection (BSI) and sepsis—a syndromic, often fatal, inflammatory response. Rapid and accurate antimicrobial prescriptions are critical to decreasing mortality in BSI patients. However, traditional antimicrobial susceptibility testing (AST) for BSI is time-consuming and tedious, leading clinicians to rely primarily on their experience when prescribing treatment.
Responding to the need for faster diagnostic tools, researchers from Shandong University, the Qingdao Institute of Bioenergy and Bioprocess Technology (QIBEBT) of the Chinese Academy of Sciences (CAS), and the Affiliated Hospital of Qingdao University have developed an integrated BSI-AST microfluidic chip for rapid AST from positive blood cultures (PBCs). Using the chip, the process from bacteria extraction to getting AST results takes less than 3.5 hours, thus promising to be a powerful new tool in managing bloodstream infections. The study was published recently in Analytical Chemistry.
"Traditional AST methods currently require at least two days to yield results following a positive blood culture. The delay in diagnosis compels the administration of empirical antibiotics, risking the aggravation of the patient's condition and fostering the emergence of antibiotic resistance," said MA Bo, PhD, professor at the Single-Cell Center at QIBEBT, co-author of the study. "Therefore, there is an urgent need for new technologies that can provide accurate and timely diagnostics and drug susceptibility testing."
Effectiveness of the BSI-AST microfluidic chip
In this study, the researchers designed a BSI-AST chip capable of extracting bacteria directly from PBCs within 10 minutes. Providing rapid AST results requires an additional three hours.
In a proof-of-concept study, the BSI-AST chip demonstrated its effectiveness by conducting direct AST on artificial PBCs containing E. coli, testing against 18 antibiotics, and producing results in less than 3.5 hours.
Moreover, the integrated chip was applied to the diagnosis of clinical PBCs, showing a categorical agreement of 93.3 percent with standard clinical methods. The reliable and rapid AST results of the chip highlight its great potential in clinical diagnosis. "In previous studies, microfluidic devices were mainly designed for purification and concentration of viable microorganisms derived from subculture or urine samples with simple composition," said Zhu Meijia, a PhD student at the Shandong University and first author of the study. "The practical utilization of these devices faced significant challenges due to the absence of on-chip complex sample preparation processes."
Xu Teng, BS, an assistant research fellow at the Single-Cell Center at QIBEBT and the study’s contributing author, said that the BSI-AST chip was a "significant advancement" since it could work directly from PBCs without the need for a subculture.
Clinical applicability of chip-based ASI test
The researchers achieved rapid extraction and enrichment of bacteria from PBCs by introducing a separator gel to the microfluidic chip for the first time. Centrifugal microfluidic enrichment technology also was central to the process. Furthermore, the chip's multiplexing analysis capability through antibiotic drying and array parallelization supports clinicians in optimizing antibiotic therapy for BSI patients.
The BSI-AST chip also provides a rapid and convenient solution for sample pretreatment when combined with Clinical Antimicrobial Susceptibility Test Ramanometry (CAST-R™), an instrument that the team invented, according to Xu Jian, PhD, professor and director of the Single-Cell Center at QIBEBT.
- This press release is supported by the Chinese Academy of Sciences