Rapid, Affordable and Portable Medium-Throughput Molecular Device for Zika Virus

Abstract

Zika virus (ZIKV) has gained global attention as an etiologic agent of fetal microcephaly and Guillain-Barré syndrome. Existing immuno-based rapid tests often fail to distinguish between Zika and related flaviviruses that are common in affected regions of Central and South Americas and the Caribbean. The US CDC and qualified state health department laboratories can perform the reverse transcription polymerase chain reaction (RT-PCR) ZIKV test using highly sophisticated instruments with long turnaround times. The preliminary results of a portable and low-cost molecular diagnostics system for ZIKV infection are reported here. In less than 15 minutes, this low-cost platform can automatically perform high quality RNA extraction from up to 12 ZIKV-spiked urine samples simultaneously. It can also perform reverse transcription recombinase polymerase amplification reaction (RT-RPA) in ≤15 minutes. The fluorescent signal produced from probe-based RT-RPA or RT-PCR assays can be monitored using LEDs and a smartphone camera. In addition, the RT-RPA and RT-PCR assays do not cross-react with dengue and chikungunya viral RNA. This low-cost system lacks complicated, sensitive and high cost components, making it suitable for resource-limited settings. It has the potential to offer simple sample-to-answer molecular diagnostics and can inform healthcare workers of patients’ diagnosis promptly.

Recombinase polymerase amplification: Basics, applications and recent advances

“…The system allows the continuous movement and heating of droplets achieving an improved detection limit (>2 orders of magnitude) as compared to benchtop assays [13]. In another report, a commercial 3D printer was modified and coupled with blue LEDs and a mobile phone camera to construct a robotic device for DNA/RNA extraction, amplification and real time detection in a multiplex format (up to 12 samples), and applied to ZIKA spiked urine samples [97]. An alternative approach combining ligation based assays with qRPA for the detection of fusion gene mRNAs was described.…”

Diagnostics in space: will zero gravity add weight to new advances?

“…Our previous study with an earlier prototype of our 3D printer-based device showed comparable extraction efficiency of C. trachomatis DNA from urine samples when compared to a gold-standard spin-column-based method from Qiagen [15]. We have also successfully extracted RNA from Zika virus samples [16]. Our recent work has focused on completing and enhancing extraction inside an enclosed setting in microgravity.…”

Involvement of the open-source community in combating the worldwide COVID-19 pandemic: a review

“…3D printed attachments allowed for dual-purposing the motors as centrifuges [63]. A modified 3D printer has been used as a gantry to perform automated allocation of samples into wells for reverse transcription PCR, using the extruder heater to heat the samples [64]. A further device has been developed for rapidly sampling air and capturing micro-organisms, which were then identified using qPCR [65].…”