Central nervous system (CNS) infections are a serious global health concern, often caused by bacterial, viral, fungal, or parasitic invasions into the brain, spinal cord, or meninges. These infections can lead to meningitis, encephalitis, myelitis, and cerebral abscesses, resulting in high mortality rates and long-term neurological disabilities. The challenge lies in the rapid and accurate identification of the pathogens responsible, as timely treatment is crucial for improved clinical outcomes.
This study aimed to develop an innovative diagnostic approach, targeted nanopore sequencing (tNPS), to address the limitations of conventional methods. tNPS offers a culture-independent, high-throughput, and efficient way to detect CNS pathogens. By utilizing specific primers for prevalent pathogens and universal primers for 16S ribosomal RNA (16S rRNA) and internal transcribed spacer (ITS) regions, tNPS expands pathogen coverage and provides a comprehensive diagnostic tool.
The study focused on 17 clinically relevant pathogens, including bacteria like Streptococcus pneumoniae and Neisseria meningitidis, fungi like Cryptococcus neoformans, and neurotropic viruses such as herpes simplex virus types 1 and 2. The tNPS method was tested on reference strains, mock communities, and clinical cerebrospinal fluid (CSF) samples.
The results demonstrated the accuracy and specificity of tNPS in detecting individual pathogens and microbial communities. The limit of detection (LOD) was determined to be 10^3 bacteria/mL, with an optimal LOD of 10^2 bacteria/mL for certain pathogens. The clinical application of tNPS showed its potential in identifying CNS infections, with a turnaround time of just 8 hours.
Nanopore sequencing technology, with its real-time capabilities and ultralong read lengths, has revolutionized pathogen detection. This study extends its application to CNS infections, highlighting its advantages. The portability of nanopore sequencing platforms offers flexibility in diagnostic settings.
While the study has shown promising results, it is important to note that RNA viruses were not included in the assay, which may limit its applicability. Further research is needed to optimize RNA workflows, increase clinical sample sizes, and conduct multicenter trials to fully evaluate the performance of tNPS.
In conclusion, the development of tNPS has the potential to revolutionize the diagnosis of CNS infections, enabling timely and accurate treatment. With its high-throughput and efficient nature, tNPS could significantly improve patient management and infection control, ultimately saving lives.
