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.

A High-Throughput, Multi-Index Isothermal Amplification Platform for Rapid Detection of 19 Types of Common Respiratory Viruses Including SARS-CoV-2

“…Among various microfluidic technologies, such as hydrophilic–hydrophobic patterning [9] , [10] , the SlipChip [11] , [12] , [13] , [14] , and electrowetting-on-dielectric (EWOD) technology [15] , [16] , the centrifugal-based method offers the advantages of simple operations, fully enclosed microstructures, and easy integration with other microfluidic functions [17] , [18] . Our group previously developed a disc-shaped centrifugal microdevice operated in a compact instrument with a single channel (only one sample tested per run) and proved the excellent multiplexing capability of this system by demonstrating the detection of 13 respiratory pathogenic bacteria using loop-mediated isothermal amplification (LAMP) on a single chip [19] . However, when viruses are detected, neither RT-PCR nor LAMP are amenable for use with a microchip due to thermal cycling and primer design problems, respectively.…”

Rapid, accurate, and novel diagnostic technique for respiratory pathogens: Clinical application of loop-mediated isothermal amplification assay in older patients with pneumonia, a multicenter prospective observational study

“…We searched PubMed database for articles with the keywords “LAMP, pneumonia or lower respiratory tract infections.” There was no language restriction, and COVID-19 was removed. We found three results ( Hou et al, 2018 ; Scharmann et al, 2020 ; Wang et al, 2020 ), all three of which collected respiratory tract specimens and submitted them for LAMP and routine clinical pathogenic testing, but did not confirm and interpret the controversial results from a clinical perspective. The indicators detected by LAMP technology are limited to 2–9 kinds of bacteria, and no attention was paid to the elderly.…”

“…20173401346). Their previous data found that compared with DNA sequencing, the sensitivity and specificity of CCID are over 95 and 99%, respectively ( Hou et al, 2018 ). However, the application value of CCID has not been verified by rigorous clinical studies.…”

Clinical features and etiology of patients with community-acquired pneumonia in southern China

“…In addition, the percentages of bacterial pathogen infection with MRSA patients have systemic disease (21.9%) was relatively higher than patients without systemic disease group (P = 0.001). MRSA is the most common bacterial pathogen infection that gives rise to a high burden of signi cant healthcare costs, morbidity, and mortality for patients every year [24][25][26]. The percentages of bacterial pathogen infection with P. aeruginosa tested positive made up 9.4% among patients with systemic disease group (P = 0.029).…”