Clinical evaluation of the loop-mediated isothermal amplification assay for the detection of common lower respiratory pathogens in patients with respiratory symptoms

Abstract

Lower respiratory tract infections (LRTIs) are a substantial public health problem and a leading cause of significant morbidity and mortality worldwide. The aim of this study was to evaluate a commercially available loop-mediated isothermal amplification (LAMP) assay for the simultaneously detection of thirteen common lower respiratory pathogens in patients with respiratory symptoms. All participants age from 1 to 101 years old were recruited from inpatient or outpatient of Meizhou People's Hospital between October 2016 and March 2018. A total of 1767 sputum samples and 88 bronchoalveolar lavage fluid samples from patients with suspected LRTI were collected. For each sample, a parallel study using both routine bacterial culture-based and LAMP assays were carried out. In total, 810 (44.85%) out of the 1855 samples were found to be positive infected with respiratory pathogens by using the LAMP assays. Methicillin-resistant Staphylococcus aureus (MecA) was the most predominant bacterial pathogens, with proportions of 17.09% in sputum and 10.23% bronchoalveolar lavage fluid samples, respectively. The proportions of bacterial pathogen infection with Streptococcus pneumoniae (Spn) (24.24%) was relatively high in aged <15 group (P <.001) while the proportions of bacterial pathogen infection with MecA (22.89%) was relatively high in aged >60 group (P <.001). Bacterial pathogen infection with MecA having the highest prevalence with proportions of 17.81% and 13.94% in male and female, respectively. A statistically higher proportion of male group had bacterial pathogen infection with Pseudomonas aeruginosa (Pae) in this study (P = .035). Comparison of results between the LAMP assay and culture method was conducted and our results indicated that there was higher detection rate by the LAMP assay than the bacterial culture method. Comparison of the results obtained with the LAMP assay and those obtained by sequencing analysis, when the sequencing method was set to 100%, demonstrating that the LAMP assay is 100% specific and 95.50% sensitive. The technique of LAMP assay was proved to be a simple, sensitive, specific, convenient, and rapid method, which can be implemented for diagnosing pathogenic bacteria in patients with LRTIs in primary labs without any need for expensive equipment or specialized techniques in resource-limited areas of China.

Figure 1

Schematic of the microfluidic chip consisting of an array of 24 reaction wells and microchannel for sample inlet, distribution and reaction. (A) schematic drawing of microfluidic chip and (B) photograph.

Figure 2

Microcentrifuge and setup CapitalBio RTisochip. (A) Microcentrifuge that used to make the mixture drop into the bottom of the reaction wells. (B) CapitalBio RTisochip-A equipped with a temperature-controlled system and a real-time imaging system. (C) Amplification curves for respiratory pathogens detection obtained using the microfluidic chip.