Clinical evaluation of probe capture-based targeted next-generation sequencing in suspected infected pancreatic necrosis: a prospective pilot diagnostic study.

OBJECTIVE

To evaluate the diagnostic performance of probe capture-based targeted next-generation sequencing (tNGS) in suspected infected pancreatic necrosis (IPN).

METHODS

A prospective study was conducted among patients with suspected IPN at Xiangya Hospital from December 2023 to May 2025. Blood samples were collected and sent for tNGS, metagenomic next-generation sequencing (mNGS), and culture. For patients with indications for surgical interventions, peripancreatic specimens were simultaneously collected for microbial culture during the initial surgical procedure to confirm the presence of IPN. The clinical performance of tNGS, mNGS and culture were compared.

RESULTS

In 51 patients, blood tNGS demonstrated a significantly higher positivity rate compared to blood culture (54.9% vs. 17.7%, P < 0.05), while no statistically significant difference was observed between blood tNGS and blood mNGS (54.9% vs. 51.0%, P = 0.683). tNGS demonstrated superior detection of fungi and Gram-negative bacteria. With peripancreatic culture as the reference standard, tNGS demonstrated superior sensitivity (94.7%), accuracy (81.3%), and concordance rate (78.9%) compared to conventional culture, and exhibited comparable performance to mNGS. tNGS also detected antimicrobial-resistance genes in 7 patients, with findings consistent with available phenotypic susceptibilities. Detection of virulence factors (VF) was associated with higher rates of persistent organ failure (POF) (100.0% vs. 57.1%, P = 0.009) and mortality (77.7% vs. 11.9%, P = 0.022). Firth’s bias-reduced logistic regression identified OF > 7 days as an independent predictor of tNGS positivity (OR 4.259, P = 0.040).

CONCLUSIONS

Blood tNGS demonstrated diagnostic performance comparable to mNGS in suspected IPN, with potentially greater sensitivity for fungal and Gram-negative pathogens. Its ability to detect VF may offer prognostic insights, but clinical utility requires further validation.