Rapid Detection of Bacterial Pathogens Causing Lower Respiratory Tract Infections via Microfluidic-Chip-Based Loop-Mediated Isothermal Amplification

Summary

Various bacterial pathogens can cause respiratory tract infections and lead to serious health issues if not detected accurately and treated promptly. Rapid and accurate detection of these pathogens via loop-mediated isothermal amplification provides effective management and control of respiratory tract infections in clinical settings.

Abstract

Respiratory tract infections (RTIs) are among the most common problems in clinical settings. Rapid and accurate identification of bacterial pathogens will provide practical guidelines for managing and treating RTIs. This study describes a method for rapidly detecting bacterial pathogens that cause respiratory tract infections via multi-channel loop-mediated isothermal amplification (LAMP). LAMP is a sensitive and specific diagnostic tool that rapidly detects bacterial nucleic acids with high accuracy and reliability. The proposed method offers a significant advantage over traditional bacterial culturing methods, which are time-consuming and often require greater sensitivity for detecting low levels of bacterial nucleic acids. This article presents representative results of K. pneumoniae infection and its multiple co-infections using LAMP to detect samples (sputum, bronchial lavage fluid, and alveolar lavage fluid) from the lower respiratory tract. In summary, the multi-channel LAMP method provides a rapid and efficient means of identifying single and multiple bacterial pathogens in clinical samples, which can help prevent the spread of bacterial pathogens and aid in the appropriate treatment of RTIs.

Introduction

Respiratory tract infections (RTIs) caused by bacterial pathogens primarily contribute to morbidity and mortality worldwide¹. It is defined as any upper or lower respiratory symptoms accompanied by fever lasting 2-3 days. While upper respiratory infection is more common than lower respiratory infection, chronic and recurrent respiratory tract infections are also common clinical conditions, posing great risks to individuals and placing a significant burden on healthcare systems². Common bacterial pathogens of RTIs include Streptococcus pneumoniae³, Haemophilus influenzae

Protocol

All samples for this study were evaluated and approved by the Ethics Review Committee of Guangdong Provincial People's Hospital (Approval Number: KY2023-1114-01). All participants signed written informed consent before the experiments. The reagents and equipment used for the study are listed in the Table of Materials. The abbreviations used in the protocol are listed in Supplementary Table 1.

1. Collection of clinical samples from the lower respirato.......

Discussion

Respiratory tract infections are prevalent hospital-associated infections, imposing severe consequences on patients and escalating mortality rates¹⁶. Timely and accurately identifying potential pathogens followed by effective antibiotics is the key to successful treatment and improving prognosis, particularly given the limitations inherent in traditional culture methods¹⁷. In this study, we used a LAMP-based method to determine single or multiple infections for fast and pre.......

Materials

Name	Company	Catalog Number	Comments
Bath Incubator(MK2000-2)	ALLSHENG		Provide a constant temperature environment
Bronchial lavage fluid collector head	TIANPINGHUACHANG	SEDA 20172081375	Collecting bronchoalveolar lavage fluid
Fiberoptic bronchoscope	OLYMPUS	SEDA 20153062703	A flexible bronchoscope equipped with a fiberoptic light source and camera, to visually examine the airways and structures within the lungs. Assist in collecting bronchoalveolar lavage
HR1500-B2	Haier	SEDA 20183541642	Biosafety cabinet
NAOH	MACKLIN	S817977	Liquefy viscous lower respiratory tract sample
Nucleic acid detection kit for respiratory tract pathogens	Capitalbio Technology	SEDA 20173401346	Testing for bacteria infection
Nucleic acid extraction reagent	Capitalbio Technology	SEDA 20160034	For DNA extraction
RTisochip-W	Capitalbio Technology	SEDA 20193220539	Loop-mediated Isothermal Amplification
THERMO ST16R	Thermo Fisher Scientific	SEDA 20180585	Centrifuge the residual liquid off the wall of the tube.
Vortex mixer VM-5005	JOANLAB		For mixing reagent