Validation of use of the miniPCR thermocycler for Ebola and Zika virus detection

Abstract

The development of point-of-care (POC) diagnostic systems has received well-deserved attention in recent years in the scientific literature, and many experimental systems show great promise in real settings. However, in the case of an epidemic emergency (or a natural disaster), the first line of response should be based on commercially available and validated resources. Here, we compare the performance and ease of use of the miniPCR, a recently commercially available compact and portable PCR device, and a conventional thermocycler for the diagnostics of viral nucleic acids. We used both thermocyclers to detect and amplify Ebola and Zika DNA sequences of different lengths (in the range of 91 to 300 nucleotides) at different concentrations (in the range of ~50 to 4.0 x 10 8 DNA copies). Our results suggest that the performance of both thermocyclers is quite similar. Moreover, the portability, ease of use, and reproducibility of the miniPCR makes it a reliable alternative for point-of-care nucleic acid detection and amplification.

Portable and accurate diagnostics for COVID-19: Combined use of the miniPCR thermocycler and a well-plate reader for SARS-CoV-2 virus detection

“…We recently published a comparison of the performance of the miniPCR and a commercial thermal cycler for the identification of artificial Zika and Ebola genetic sequences. Our experiments using a wide variety of primers sets and template concentrations revealed no differences in performance between either thermal cycler type [37]. The commercial availability, low price (as compared to conventional thermocyclers), portability, and user friendliness of the miniPCR makes it an attractive and tangible solution that effectively brings PCR analysis to the POC.…”

“…Time is the most limiting factor in epidemic emergencies. Therefore, the integration of well-developed and commercially available technologies [ 31 , 37 , 38 ] becomes an obvious, expedient, and cost-effective first line of defense in the context of COVID-19 pandemics. Here, we demonstrate that the combined use of a commercial and portable PCR unit (the miniPCR) and a 96-well plate reader is potentially adequate for the fast deployment of diagnostic efforts.…”

“…We used a plasmid containing the GP gene from Ebola Virus (EBOV) as a negative control. The production of this EBOV genetic material has been documented previously by our group [37].…”

“…Only recently, several miniaturized PCR machines become commercially available [ 28 , 29 ]. One of them, the miniPCR [ 30 , 31 ], reached the international market in 2015. The most recent version of this compact PCR machine has an approximate cost of ~$800 USD ( www.minipcr.com ) as compared to $3000 USD for a conventional PCR thermocycler [ 28 ].…”

Diagnostic validation of a rapid and field-applicable PCR-lateral flow test system for point-of-care detection of cyprinid herpesvirus 3 (CyHV-3)

“…However, many of these arguments can be discussed controversially. Today, low-cost thermal cyclers suitable for POC analyses are available, which generally allow PCR-based rapid and “easy-to-handle” field applications [ 36 , 37 ]. Further improvement of the PCR components can simplify PCR processing significantly (e.g.…”