Predicting Viruses Accurately by a Multiplex Microfluidic Loop-Mediated Isothermal Amplification Chip

Abstract

Multiplex gene assay is a valuable molecular tool not only in academic science but also in clinical diagnostics. Multiplex PCR assays, DNA microarrays, and various nanotechnology-based methods are examples of major techniques developed for analyzing multiple genes; none of these, however, are suitable for point-of-care diagnostics, especially in resource-limited settings. In this report, we describe an octopus-like multiplex microfluidic loop-mediated isothermal amplification (mμLAMP) assay for the rapid analysis of multiple genes in the point-of-care format and provide a robust approach for predicting viruses. This assay with the ability of analyzing multiple genes qualitatively and quantitatively is highly specific, operationally simple, and cost/time-effective with the detection limit of less than 10 copies/μL in 2 μL quantities of sample within 0.5 h. We successfully developed a mμLAMP chip for differentiating three human influenza A substrains and identifying eight important swine viruses.

Microfluidic method for rapid turbidimetric detection of the DNA of Mycobacterium tuberculosis using loop-mediated isothermal amplification in capillary tubes

“…These micro-reactors utilized the rapid thermal conduction from increased surface-to-volume ratio, as well as the low sample/reagent consumption attributable to reduce the size of the container [25][26][27]. For this purpose, various approaches of microfluidic LAMPs were reported for indicating the superiority in terms of the speed, the miniaturization, the power consumption, and the cost beneficence [28][29][30][31]. Capillary tubes could also be applied for the microfluidic-LAMP reaction instead of some polymeric materials (e.g.…”

High-throughput sample-to-answer detection of DNA/RNA in crude samples within functionalized micro-pipette tips

“…Nucleic acid (deoxyribonucleic acid, DNA and ribonucleic acid, RNA) analysis is emerging as one of the most effective methods in biomedical applications Fang et al 2012;Fang et al 2010a;Fang et al 2010b;Liu et al 2011;Sun et al 2014;Wu et al 2015;Xie et al 2015;Zhang et al 2014a;Zhang et al 2014b), especially for diagnosis of infectious diseases.…”

“…Despite of well-established approaches for nucleic acid tests (NAT) such as polymerase chain reaction (PCR) (Urdea et al 2006), these methods may not be affordable or accessible in resource-poor settings due to the lack of modern healthcare infrastructure, such as laboratory of high biosafety level and expensive PCR instruments (Fang et al 2010a;Fang et al 2010b;Urdea et al 2006;Zuo et al 2013). In fact, the prevalence of infectious diseases in developing countries is much higher than that in developed world (Lee et al 2010;Peeling and Mabey 2010).…”

l-cysteine induced manganese porphyrin electrocatalytic amplification with 3D DNA-Au@Pt nanoparticles as nanocarriers for sensitive electrochemical aptasensor

“…Traditional methods for immobilizing signal tags and enzymes include hybridization chain reaction (HCR) Zhang et al, 2012), rolling circle amplification (RCA) Li et al, 2011), loop-mediated isothermal amplification (LAMP) (Wang et al, 2010;Fang et al, 2011), and strand displacement amplification (SDA) (Zhang et al, 2013;Weizmann et al, 2006). Recently, a novel DNA based 3D structure formed by the hybridization reaction of two single DNA strands has be invented Sun et al, 2011).…”