The U.S. Army Contracting Command – Aberdeen Proving Ground (ACC-APG), Natick Contracting Division is conducting market research to obtain information regarding the capability of companies to manufacture Ebola Zaire Real-Time PCR (EZ rRT-PCR) assays which includes all the primers, probes, and reaction buffers.
The DoD-developed “EZ1” Ebola diagnostic test was approved for emergency use by the Food and Drug Administration (FDA) in Oct. 2014.
The validated and approved platforms for the assay to be run on are the Applied Biosystems 7500 Fast Dx Real-Time PCR Instrument (ABI 7500 Fast Dx), Roche LightCycler (LightCycler), and Biofire Defense Joint Biological Agent Identification and Diagnostic System (JBAIDS).
The level of Ebola Zaire virus will be detected with the EZ1 assay via presence in blood from individuals with early systemic infection. The definitive identification of the Ebola Zaire Virus will require additional testing and confirmation procedures in consultation with public health or other authorities for whom reporting is required.
The EZ1 assay requires the following reagents:
- EZ1 Master Mix – Contains primer and probe sequences that specifically detect Ebola Zaire in Trizol-inactivated whole blood or Trizol-inactivated plasma specimens.
- RNase P (RP) – Used to detect human RP and is used as a positive control with human clinical specimens to indicate that adequate isolation of nucleic acid resulted from the extraction of the clinical specimen.
- EZ1 Positive Template Control (EZ1-PTC) – This control will be PTC designed to react with the EZ1-PCR reagents to indicate whether the Ebola Zaire PCR reaction worked. This PTC material consists of synthetic template RNA.
- RP Positive Template Control (RP-PTC) – Designed to react with the RP rRT-PCR reagents to indicate whether the RP PCR reaction worked. This PTC material consists of synthetic template RNA. The RP-PTC should only be positive for the RP assay.
- RT- Taq Polymerase – This must be the SuperScript One-Step RT-PCR System with Platinum Taq DNA Polymerase sold by Thermo Fisher.
The government states that the SuperScript consumable must remain as part of the manufactured assays, as it was included in the original Emergency Use Authorization submission to the FDA. As such, to establish an additional PCR production facility, the manufacturer would need to obtain licensing for the intellectual property (IP) associated with SuperScript product which is owned by Roche, Ltd. In order to change the consumable, the FDA requires proof of performance through equivalency testing with changed variable.
Additional details are available under Solicitation Number: W911QY-15-S-0001.
