Mobile and accurate detection system for infection by the 2009 pandemic influenza A (H1N1) virus with a pocket-warmer reverse-transcriptase loop-mediated isothermal amplification

Abstract

The 2009 pandemic H1N1 influenza A virus spread quickly worldwide in 2009. Since most of the fatal cases were reported in developing countries, rapid and accurate diagnosis methods that are usable in poorly equipped laboratories are necessary. In this study, a mobile detection system for the 2009 H1N1 influenza A virus was developed using a reverse-transcriptase loop-mediated isothermal amplification (RT-LAMP) kit with a disposable pocket-warmer as a heating device (designated as pwRT-LAMP). The pwRT-LAMP can detect as few as 100 copies of the virus--which is nearly as sensitive as real-time reverse-transcription polymerase chain reaction (RT-PCR)--and does not cross-react with RNA of seasonal influenza viruses. To evaluate the usefulness of the pwRT-LAMP system, nasal swab samples were collected from 56 patients with flu-like symptoms and were tested. Real-time RT-PCR confirmed that the 2009 H1N1 influenza A virus was present in 27 of the 56 samples. Of these 27 positive samples, QuickVue Influenza A+B immunochromatography detected the virus in only 11 samples (11/27; 40.7%), whereas the pwRT-LAMP system detected the virus in 26 of the 56 samples (26/27 of the positive samples; 96.3%). These findings indicate that the mobile pwRT-LAMP system is an accurate diagnostic system for the 2009 H1N1 influenza A virus, and has great potential utility in diagnosing future influenza pandemics.

Emerging Loop-Mediated Isothermal Amplification-Based Microchip and Microdevice Technologies for Nucleic Acid Detection

“…In a similar approach, an electricity-free cartridge was integrated with LFT for HIV detection with a sensitivity of 6 copies/ μ L. 94 It was also shown that even a pocket hand warmer could be used for LAMP amplification. 95 Zhang et al 96 used a pocket warmer and capillary glass for LAMP-based multiplexing target NAs. Various target DNAs and negative controls were injected at different positions of the channel, and the two sides of the channel were sealed with epoxy glue.…”

Two Years after Pandemic Influenza A/2009/H1N1: What Have We Learned?

“…The most useful laboratory test for the clinical management of A/2009/H1N1 influenza is nucleic acid amplification after reverse transcription (Table 11). In 10 studies, the overall reported sensitivity of real-time RT-PCR ranged from 90.5% to 97.6% (60,202,240,286,321,336,404,422,432,559). The majority of the studies reported a specificity of 100%, with four exceptions (60,404,432,559).…”

Specific Detection of Klebsiella variicola and K. oxytoca by Loop-Mediated Isothermal Amplification

“…As a result, this technology has been used widely for molecular detection of several microorganisms, including clinical and plant-associated bacterial [32,[61][62][63][64][65][66][67][68][69][70][71][72][73], fungal [74][75][76], viral [47,[77][78][79][80][81][82][83][84], and parasitic [57,59,[85][86][87][88] pathogens, and is therefore suitable as a rapid field test.…”