OPEN Analytical chemistry | 17 Jun 2016
J Song, MG Mauk, BA Hackett, S Cherry, HH Bau and C Liu
The recent outbreak of Zika virus (ZIKV) infection in the Americas and its devastating impact on fetal development have prompted WHO to declare the ZIKV pandemic as a Public Health Emergency of International Concern. Rapid and reliable diagnostics for ZIKV are vital since ZIKV-infected individuals display no symptoms or nonspecific symptoms similar to other viral infections. Since 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 on the assay 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 any electrical power. Detection is carried out with leuco crystal violet (LCV) dye by eye, thus eliminating the need for any 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 and at home.
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