Isolation ofFusobacterium nucleatumfrom human feces using immunomagnetic separation coupled with fastidious anaerobe agar

Abstract

AbstractAimFusobacterium nucleatum (F. nucleatum) is associated with the initiation, development, and metastasis of colorectal cancer. However, it is difficult to isolate F. nucleatum from clinical specimens. In this study, we aimed to develop an effective and rapid method for isolating F. nucleatum from human feces using polyclonal antibody (PAB)-coated immunomagnetic beads (IMBs) with selective media.Methods and resultsIMBs conjugated with PAB were prepared and used to isolate F. nucleatum from human feces, and the bacteria were cultured with selective culture media (fastidious anaerobe agar + nalidixic acid + vancomycin). Under optimized experimental conditions, IMBs could selectively recover F. nucleatum from fecal microbiota samples spiked with Peptostreptococcus or Bacteroides fragilis. In artificial fecal samples, the detection sensitivity of IMBs for F. nucleatum was 103 CFU mL−1. In addition, IMBs combined with selective media could rapidly isolate F. nucleatum from human feces.ConclusionsThis study successfully established an effective method for the rapid isolation of F. nucleatum from human feces by IMBs. The whole procedure requires 2–3 days, and has a sensitivity of 103 CFU mL−1 feces.