Evaluation of Nucleic Acid Isothermal Amplification Methods for Human Clinical Microbial Infection Detection

Area of Science:

• Medical Diagnostics
• Molecular Biology
• Infectious Diseases

Background:

• Prompt identification of infection is crucial for effective treatment and preventing sepsis.
• Current pathogen identification methods are time-consuming, often requiring days or extensive laboratory procedures.
• Broad-spectrum empirical therapy is frequently used due to delays in precise pathogen detection.

Purpose of the Study:

• To develop and evaluate a rapid genetic diagnostic method for identifying common infection pathogens.
• To assess the performance of the Infection Diagnosis Panel (In-Dx) across various clinical sample types.
• To enable faster and more precise diagnosis for point-of-care utilization.

Main Methods:

• Utilized loop-mediated isothermal amplification (LAMP) for rapid (70-120 min) genetic detection of 15 common infection pathogens.
• Employed filtration for bacterial concentration and direct LAMP amplification without DNA purification.
• Tested the In-Dx panel on clinical samples (blood, urine, wound, sputum, stool) using fluorescent and visual colorimetric detection.

Main Results:

• The In-Dx panel demonstrated high sensitivity and specificity for pathogen detection in urine (Se: 91.1%, Sp: 97.3%) and mucocutaneous swabs (Se: 65.5%, Sp: 99.3%).
• Achieved excellent negative predictive values (NPV) for both sample types (Urine: 99.7%, Swabs: 98%).
• The method successfully identified targeted pathogens from diverse clinical matrices.

Conclusions:

• The LAMP-based In-Dx panel provides a rapid and precise method for diagnosing clinical infections.
• This diagnostic approach supports timely antimicrobial treatment decisions.
• The In-Dx panel is suitable for point-of-care testing across multiple sample types.