Targeted next-generation sequencing enhances precision and rapid detection in healthcare-associated infection Surveillance: Unveiling multidrug-resistant colonization in ICUs.

Objectives: This study aims to evaluate the potential advantages of targeted next-generation sequencing (tNGS) over conventional bacterial culture methods for pathogen detection in hospital-associated infections (HAIs).

Methods: All EICU medical staff and all medical staff from the Physical Examination Centre completed a questionnaire. Nasopharyngeal specimens were collected from medical staff who met all of the inclusion criteria and none of the exclusion criteria. EICU medical staff provided 2 samples each, while Physical Examination Centre staff provided 1 sample each. For EICU medical staff, one of their two nasopharyngeal swabs was subjected to tNGS testing, and the other to bacterial culture testing. For the PEC staff, their nasopharyngeal swabs were subjected to tNGS testing. Additionally, six pairs of spectacles and six keyboards used by EICU medical staff were randomly selected, and the surfaces were swabbed with sterile swabs for tNGS testing.

Results: In 23 nasal swab samples from EICU group, tNGS detected 14 species of microorganism in 29 instances within 19 h. Bacterial culture detected 2 species of microorganism in 4 instances, 2 positive samples within 19 h and confirmed another 2 positive samples within 69 h. A total of 42 samples with 14 different microorganism species were collected from the nasopharyngeal swabs of 23 EICU members and 15 PEC members. Among them, 29 cases (69 %) of 14 different microorganisms were detected in EICU staff, with an average of 1.3 microorganism species detected per person, while 13 cases (28 %) of 6 different microorganisms were detected in PEC staff, with an average of 0.9 microorganism species detected per person. The most common colonizing bacteria included , , and spp. Compared to bacterial culture, tNGS offers advantages in monitoring HAIs, including a broad range of detectable microorganisms, high sensitivity of results, and shorter reporting time for positive results. Bacteria colonizing the EICU carry more antibiotic resistance genes.

Conclusions: tNGS outperforms conventional culture in healthcare-associated infection surveillance, with higher sensitivity and accelerated pathogen identification. Simultaneously, tNGS revealed extensive colonization of multidrug-resistant (MDR) pathogens (e.g., , MRSA) in EICU environments, highlighting its utility in monitoring complex antimicrobial resistance patterns.