title: Multiplexed instrument-free meningitis diagnosis on a polymer/paper hybrid microfluidic biochip.
abstract: Neisseria meningitidis (N. meningitidis), Streptococcus pneumoniae (S. pneumoniae), and Haemophilus influenzae type b (Hib) are three most common pathogens accounting for most bacterial meningitis, a serious global infectious disease with high fatality, especially in developing nations. Because the treatment and antibiotics differ among each type, the identification of the exact bacteria causing the disease is vital. Herein, we report a polymer/paper hybrid microfluidic biochip integrated with loop-mediated isothermal amplification (LAMP) for multiplexed instrument-free diagnosis of these three major types of bacterial meningitis, with high sensitivity and specificity. Results can be visually observed by the naked eye or imaged by a smartphone camera under a portable UV light source. Without using any specialized laboratory instrument, the limits of detection of a few DNA copies per LAMP zone for N. meningitidis, S. pneumoniae and Hib were achieved within 1h. In addition, these three types of microorganisms spiked in artificial cerebrospinal fluid (ACSF) were directly detected simultaneously, avoiding cumbersome sample preparation procedures in conventional methods. Compared with the paper-free non-hybrid microfluidic biochip over a period of three months, the hybrid microfluidic biochip was found to have a much longer shelf life. Hence, this rapid, instrument-free and highly sensitive microfluidic approach has great potential for point-of-care (POC) diagnosis of multiple infectious diseases simultaneously, especially in resource-limited settings.
authors.affiliation: Department of Chemistry, University of Texas at El Paso, 500 West University Ave, El Paso, TX 79968, USA.
authors.forename: Maowei
authors.initials: M
authors.lastname: Dou
authors.affiliation: Department of Chemistry, University of Texas at El Paso, 500 West University Ave, El Paso, TX 79968, USA.
authors.forename: Sharma T
authors.initials: ST
authors.lastname: Sanjay
authors.affiliation: College of Health Sciences, University of Texas at El Paso, 500 West University Ave, El Paso, TX 79968, USA.
authors.forename: Delfina C
authors.initials: DC
authors.lastname: Dominguez
authors.affiliation: Department of Biomedical Engineering, School of Medicine, Collaborative Innovation Center for Diagnosis and Treatment of Infectious Diseases, Tsinghua University, Beijing, 100084, China.
authors.forename: Peng
authors.initials: P
authors.lastname: Liu
authors.affiliation: Bioinspired Engineering and Biomechanics Center (BEBC), Xi'an Jiaotong University, Xi'an 710049, PR China.
authors.forename: Feng
authors.initials: F
authors.lastname: Xu
authors.affiliation: Department of Chemistry, University of Texas at El Paso, 500 West University Ave, El Paso, TX 79968, USA; Biomedical Engineering, University of Texas at El Paso, 500 West University Ave, El Paso, TX 79968, USA; Border Biomedical Research Center, University of Texas at El Paso, 500 West University Ave, El Paso, TX 79968, USA; Environmental Science and Engineering, University of Texas at El Paso, 500 West University Ave, El Paso, TX 79968, USA. Electronic address: xli4@utep.edu.
authors.forename: XiuJun
authors.initials: X
authors.lastname: Li
mesh_terms: D015374:Biosensing Techniques / Q000295:instrumentation*; D004269:DNA, Bacterial / Q000032:analysis / Q000302:isolation & purification; D004867:Equipment Design; D006192:Haemophilus Infections / Q000134:cerebrospinal fluid / Q000175:diagnosis / Q000382:microbiology; D006193:Haemophilus influenzae / Q000302:isolation & purification*; D006801:Humans; D056656:Lab-On-A-Chip Devices; D057230:Limit of Detection; D008585:Meningitis, Meningococcal / Q000134:cerebrospinal fluid / Q000175:diagnosis* / Q000382:microbiology; D009345:Neisseria meningitidis / Q000302:isolation & purification*; D010209:Paper; D011008:Pneumococcal Infections / Q000134:cerebrospinal fluid / Q000175:diagnosis / Q000382:microbiology; D019095:Point-of-Care Systems; D011108:Polymers / Q000737:chemistry; D013296:Streptococcus pneumoniae / Q000302:isolation & purification*
chemical_list: D004269:DNA, Bacterial; D011108:Polymers
keywords: Hybrid microfluidic biochip; Instrument-free detection; LAMP; Low-resource settings; Multiplexed meningitis diagnosis