Streamlining Borrelia burgdorferi cultivation using quantitative PCR screening

Area of Science:

• Microbiology
• Infectious Diseases
• Molecular Diagnostics

Background:

• Borrelia burgdorferi culture is the gold standard for Lyme disease confirmation but is rarely used due to low success rates and lengthy protocols.
• Direct detection of B. burgdorferi is crucial for accurate Lyme disease diagnosis and management.

Purpose of the Study:

• To streamline Borrelia burgdorferi culture by integrating quantitative PCR (qPCR) screening.
• To identify cultures likely to yield viable spirochetes early in the process.

Main Methods:

• Blood plasma and cerebrospinal fluid (CSF) samples from children with confirmed Lyme disease were cultured for up to 9 weeks.
• Quantitative PCR (qPCR) targeting the B. burgdorferi flagellin B gene was used to assess DNA presence at week 3.
• qPCR results were compared to the assay's 95% limit of detection (LOD).

Main Results:

• Viable spirochetes were observed in 6.3% of plasma cultures and 27.3% of CSF cultures after 9 weeks.
• Successful cultures showed B. burgdorferi DNA copy numbers above the 95% LOD in week 3 qPCR testing.
• 100% of successful CSF cultures and 66.7% of successful plasma cultures met this qPCR criterion.

Conclusions:

• Borrelia burgdorferi culture is challenging and often unsuccessful, despite significant workload.
• qPCR serves as an effective screening tool to prioritize cultures with a higher likelihood of yielding viable spirochetes.
• Integrating qPCR screening can streamline the B. burgdorferi culturing process for improved diagnostic efficiency.