Instrument-Free Point-of-Care Molecular Detection of Zika Virus.
The recent outbreak of Zika virus (ZIKV) infection in the Americas and its devastating impact on fetal development have prompted the World Health Organization (WHO) to declare the ZIKV pandemic as a Public Health Emergency of International Concern. Rapid and reliable diagnostics for ZIKV are vital because ZIKV-infected individuals display no symptoms or nonspecific symptoms similar to other viral infections. Because immunoassays lack adequate sensitivity and selectivity and are unable to identify active state of infection, molecular diagnostics are an effective means to detect ZIKV soon after infection and throughout pregnancy. We report on a highly sensitive reverse-transcription loop-mediated, isothermal amplification (RT-LAMP) assay for rapid detection of ZIKV and its implementation in a simple, easy-to-use, inexpensive, point-of-care (POC) disposable cassette that carries out all the unit operations from sample introduction to detection. For thermal control of the cassette, we use a chemically heated cup without a need for electrical power. Amplification products are detected with leuco crystal violet (LCV) dye by eye without a need for instrumentation. We demonstrated the utility of our POC diagnostic system by detecting ZIKV in oral samples with sensitivity of 5 plaque-forming units (PFU) in less than 40 min. Our system is particularly suitable for resource-poor settings, where centralized laboratory facilities, funds, and trained personnel are in short supply, and for use in doctors' offices, clinics, and at home.
Department of Mechanical Engineering and Applied Mechanics, University of Pennsylvania , Philadelphia, Pennsylvania 19104, United States.
Jinzhao
J
Song
Department of Mechanical Engineering and Applied Mechanics, University of Pennsylvania , Philadelphia, Pennsylvania 19104, United States.
Michael G
MG
Mauk
Department of Microbiology, Perelman School of Medicine, University of Pennsylvania , Philadelphia, Pennsylvania 19104, United States.
Brent A
BA
Hackett
Department of Microbiology, Perelman School of Medicine, University of Pennsylvania , Philadelphia, Pennsylvania 19104, United States.
Sara
S
Cherry
Department of Mechanical Engineering and Applied Mechanics, University of Pennsylvania , Philadelphia, Pennsylvania 19104, United States.
Haim H
HH
Bau
Department of Mechanical Engineering and Applied Mechanics, University of Pennsylvania , Philadelphia, Pennsylvania 19104, United States.
Changchun
C
Liu
D017931:DNA Primers / Q000378:metabolism; D005840:Gentian Violet / Q000737:chemistry; D006801:Humans; D044085:Microfluidics; D021141:Nucleic Acid Amplification Techniques; D019095:Point-of-Care Systems; D012367:RNA, Viral / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D012463:Saliva / Q000821:virology; D000071244:Zika Virus / Q000235:genetics / Q000302:isolation & purification*; D000071243:Zika Virus Infection / Q000175:diagnosis / Q000821:virology
D017931:DNA Primers; D012367:RNA, Viral; D005840:Gentian Violet