Objective To compare the detection efficacy of Multiple Nucleotide Polymorphism (MNP) marker method and Microfluidic Technology for pathogens causing Severe Acute Respiratory Infections (SARI).

Methods A total of 32 respiratory specimens from SARI cases were collected from three hospitals in Ma'anshan City between March and April 2024. Pathogens were detected using both the MNP marker method and the Microfluidic method, and 14 pathogens were verified using single/dual fluorescent PCR. Statistical analysis was performed using SPSS 20.0.

Results The Microfluidic method detected 19 pathogens, with Haemophilus influenzae (68.8%), Epstein-Barr virus (28.1%), and Human Rhinovirus (12.5%) showing higher detection rates. The MNP marker method detected 24 pathogens, with Haemophilus influenzae (53.1%), Epstein-Barr virus (40.6%), and Human Herpesvirus 7 (40.6%) being the main detected pathogens. The detection results of the two methods for 24 pathogens, including SARS-CoV-2, were completely consistent (Kappa = 1.000). The consistency for Influenza A virus was good (Kappa = 0.840), while the consistency for 7 pathogens including Epstein-Barr virus was moderate (Kappa = 0.467～0.728), and the consistency for 6 pathogens including Human Rhinovirus was poor (Kappa = -0.049～0.351). The sensitivity (92.9% vs. 78.6%) and positive predictive value (100% vs. 91.7%) of the MNP marker method were superior to those of the Microfluidic method. Conclusion The MNP marker method has higher sensitivity and accuracy in detecting SARI pathogens, while the microfluidic method still requires optimization for distinguishing some closely related pathogens.