Diseases that are associated with increased body temperature (fever more than ≥ 38 °C) and other clinical signs like headache, tiredness and muscle pain are many. They can be caused by different microorganisms that are widespread in different regions, including Kenya. Sometimes these diseases can occur as outbreaks that make many people ill. One of the big problems with these types of diseases is determining the cause of the disease because symptoms such as high temperature, headache, tiredness and muscle pain are common signs of many diseases. A machine that can quickly and correctly identify microorganisms in blood of patients with elevated temperature would make treatment decisions easier for doctors. The FilmArray Global Fever (GF) Panel has the potential to detect many of these diseases. It is a machine that works by multiplying genetic material of an organism, making such materials easier to detect. The system is able to detect bacteria, viruses, and protozoa in a single blood sample. Unlike other methods that require human intervention, the machine performs all manipulations including extraction of genetic materials, multiplication of genetic materials, and identification of microorganism. The process is fully automated and gives final results in approximately one hour. The purpose of this study is to test how good this machine is in detecting the following microorganisms in patients with fever: Leptospira spp., Plasmodium spp., Plasmodium falciparum, Plasmodium vivax/ovale, chikungunya virus, and dengue virus. To do this, the machine will be tested using blood samples that will come from an ongoing study that collects blood samples from patients with fever (KEMRI SSC #1282 and WRAIR #1402). The FilmArray test results of Detected or Not Detected will be compared to validated molecular-based methods that are performed at BioFire Defense, LLC in Salt Lake City, Utah, US. Data obtained in this study will be used to support application to license the FilmArray GF Panel as an in vitro diagnostic (IVD) device for detection of microorganisms in blood from patients with fever. If the proposed study is successful, the FilmArray GF Fever Panel may provide an effective way to obtain a quick diagnosis. The study is sponsored by a company called BioFire Defense, LLC and the study will be performed at the Basic Science Lab of the Kenya Medical Research Institute/ Walter Reed Project (KEMRI/WRP) in Kisumu.

This study, entitled Clinical Evaluation of the FilmArray Global Fever (GF) Panel, is sponsored by BioFire Defense, LLC and aims to evaluate the clinical sensitivity and specificity of the assays comprising the FilmArray Global Fever Panel when used to test whole blood collected from individuals with acute febrile illness, when compared to a well-validated molecular-based method, in the intended use setting (reference or diagnostic laboratory). The intention is to use these performance data to support classification of the FilmArray GF Panel as an in vitro diagnostic (IVD) device by the U.S. Food and Drug Administration (FDA) and other comparable regulatory agencies. Whole blood will be collected prospectively from patients attending outpatient and inpatient departments of the health care facilities at Kisumu District Hospital, Kombewa Sub-District Hospital, Kisii District Hospital, Marigat District Hospital, Gilgil District Hospital, and Kenya Armed Forces Memorial Hospital (Mbagathi, Nairobi), where only enrollment and specimen collection will be performed. The subjects will be consented for blood draw and testing of pathogens that are commonly associated with febrile illnesses such as malaria, leishmaniasis, anthrax, leptospirosis, plague, paratyphoid fever, typhoid fever, tularemia, Crimean-Congo hemorrhagic fever, chikungunya fever, dengue fever, lassa fever, ebola virus disease, marburg virus disease, West Nile fever and Yellow fever. The collected blood sample will be labeled with only a study code number (SCN) in order to protect subject identity. However, in the event a select agent is detected twice using the Global Fever Panel, the enrollment log which links the SCN to the subject may be used to identify the subject for follow-up. The samples will be secured in refrigerators and freezers at WRP/KEMRI facilities and only study investigators and their authorized staff will have access. All safeguards ensuring privacy that are in place during this study period will also continue to be in place for the long-term storage of samples. Some tests planned for this trial will be done with our collaborating laboratories abroad. Before shipment of samples to such collaborating laboratories, authority for sample shipment and testing will be sought from KEMRI SERU. The collected specimens will either be tested fresh or will be frozen and then tested at a later date. Freezing of specimens will allow for continued collection of specimens when immediate testing is not possible. Freezing and testing of specimens will be tracked on study-specific forms. Informed consent will be obtained from eligible participants prior to specimen collection. For the study, up to 500 patients will provide 2.5-3.0 mL of whole blood and the blood sample will be labeled with a study code number (SCN). Continuous enrollment and testing are expected to last for approximately 18 months in order to achieve a required number of valid specimens. All FilmArray testing and nucleic acid extraction will occur at the Basic Science Lab, KEMRI/WRP, Kisumu. Of the 2.5-3.0 mL of whole blood collected, 500 L will be archived at ≤-70°C immediately for discrepancy testing. A 200 L aliquot of the whole blood will be used for FilmArray GF Panel testing. In addition to FilmArray testing, 800 L of the whole blood will be used to purify nucleic acid, using the provided MagNA Pure Compact system (or equivalent). The purified nucleic acids will be shipped frozen to BioFire Defense for further testing (comparator testing). Comparator assays are designed to amplify and detect pathogen nucleic acid; no genetic testing or specific amplification of human nucleic acid will be performed. The residual blood specimen will be stored refrigerated until FilmArray testing and nucleic acid extraction is complete in the event that additional testing/extraction is required. Once the required testing is completed, remaining sample (up to 2.0 mL total, which includes the 500 µl aliquot archived for discrepancy testing), will be archived at ≤-70°C for at least two years after study completion. In order to conduct future research utilizing these samples, a new sub-protocol must be written, reviewed and approved by both the WRAIR IRB and the KEMRI SERU. To maintain patient confidentiality, blood samples, blood sample aliquots and specimen nucleic acid will only be labeled with a study code number (SCN). This study will be performed according to the approved multisite protocol (attached), laboratory-specific protocols and site-specific workflow procedures, and available product literature for the FilmArray instrument and the MagNA Pure Compact system. No deviation from this protocol will be implemented without the prior review and approval of BioFire Defense and the respective sites’ IRB(s), except where it may be necessary to eliminate an immediate hazard to a research subject. The results of the FilmArray GF Panel testing will not be provided to the patients or their healthcare providers, or in any way be used to affect their standard of care except in the event that a true positive select agent detection is confirmed (see 3.4 below for full description). This study is jointly funded by the U.S. Army Medical Materiel Development Activity (USAMMDA) through contracts with BioFire Defense and managed by the Joint Program Executive Office for Chemical and Biological Defense – Medical Countermeasure Systems (JPEO-CBD-MCS), the National Institute of Allergy and Infectious Diseases (NIAID) and BioFire Defense. The clinical evaluation outlined in this protocol will be administered and managed by BioFire Defense. The Sponsor contact person(s) for this study will be at BioFire Defense, and are listed above (page i).