Diagnostic performance of metagenomic next-generation sequencing and conventional microbial culture for spinal infection: a retrospective comparative study

Eur Spine J. 2023 Dec;32(12):4238-4245. doi: 10.1007/s00586-023-07928-6. Epub 2023 Sep 9.

Abstract

Purpose: The study evaluated the diagnostic performance of metagenomic next-generation sequencing (mNGS) as a diagnostic test for biopsy samples from patients with suspected spinal infection (SI) and compared the diagnostic performance of mNGS with that of microbial culture.

Methods: All patients diagnosed with clinical suspicion of SI were enrolled, and data were collected through a retrospective chart review of patient records. Biopsy specimens obtained from each patient were tested via mNGS and microbial culture. Samples were enriched for microbial DNA using the universal DNA extraction kit, whole-genome amplified, and sequenced using MGISEQ-200 instrument. After Low-quality reads removed, the remaining sequences for microbial content were analyzed and aligned using SNAP and kraken2 tools.

Results: A total of 39 patients (19 men and 20 women) were deemed suitable for enrollment. The detection rate for pathogens of mNGS was 71.8% (28/39), which was significantly higher than that of microbial culture (23.1%, p = 0.016). Mycobacterium tuberculosis complex was the most frequently isolated. Using pathologic test as the standard reference for SI, thirty-one cases were classified as infected, and eight cases were considered aseptic. The sensitivity and specificity values for detecting pathogens with mNGS were 87.1% and 87.5%, while these rates were 25.8% and 87.5% with conventional culture. mNGS was able to detect 88.9% (8/9) of pathogens identified by conventional culture, with a genus-level sensitivity of 100% (8/8) and a species-level sensitivity of 87.5% (7/8).

Conclusion: The present work suggests that mNGS might be superior to microbial culture for detecting SI pathogens.

Keywords: Etiological diagnosis; Metagenomics; Molecular diagnostics; Next-generation sequencing; Spinal infection.

Publication types

  • Research Support, Non-U.S. Gov't
  • Comment

MeSH terms

  • Affect*
  • DNA
  • Female
  • High-Throughput Nucleotide Sequencing*
  • Humans
  • Male
  • Retrospective Studies
  • Sensitivity and Specificity

Substances

  • DNA