Eliminating Malaria from Haiti. Reinventing DNA to Eradicate Endemic Parasites

Eliminating Malaria from Haiti Reinventing DNA to Eradicate Endemic Parasites Firebird Biomolecular Sciences LLC University of Florida Steven A Benner John B Dame Abstract Eradication of Plasmodium falciparum the parasite that causes the most malarial deaths is the only way to finally treat the disease in any specific geographical region Haiti is one such region where a small population million geographical isolation and special ecology make eradication a real possibility Thus the Gates Foundation has just committed $ MM to the CDC Foundation to attempt this eradication Key to eradication is an assay that identifies human carriers asymptomatic individuals with active parasite infections That assay must a Require only a small sample of blood perhaps L b Detect very small numbers of living parasites perhaps as few as parasites per sample c Be as simple to operate as a conventional malarial rapid diagnostics test RDT and d cost less than $ to run These specifications are virtually impossible with classical molecular diagnostics However Firebird has developed many innovations over the past three years that make such specs possible including a sample prep processes that remove biohazard b whole nucleic acid capture without centrifugation c isothermal nucleic acid amplification using self avoiding molecular recognition systems SAMRS and artifi cially expanded genetic information systems AEGIS and d AEGIS molecular beacons Firebird has shown that these support assays that for example detect mosquito borne RNA viruses in one mosquito carcass The work is guided by work in the Dame lab showing that P falciparum ribosomal RNA present at copies per parasite can be detected in malaria infected blood using RT PCR The PCR assay does not meet the cost specs necessary for a LRE and is not as easy to run as an RDT but it shows the sensitivity of an assay directed at falciparum rRNA where detection is robust if a sample of blood contains just one organism This project will combine these innovations to develop an assay that detects falciparum rRNA as easily as an RDT but which much higher sensitivity One strength of the approach is its use of realistic samples of live falciparum in real blood to do benchmarking Costing less than $ the assay will generate fluorescence if falciparum is present that can be read directly or transmitted by cell phone camera for remote confirmation We will benchmark i a sample preparation work flow that releases P falciparum rRNA that is ii captured on a solid support where iii captured rRNA is amplified isothermally using primers with SAMRS and AEGIS nucleotides with iv amplicons detected by AEGIS beacons Results will be compared to samples analyzed by RT PCR Metrics for success include an ability to detect parasites in a sample In Phase the efficacy of the test will be validated in a small field trial with blood from individuals from in a malaria endemic area of Haiti again comparing with RT PCR This will as well support the epidemiological science behind carrier identifica tion and eradication as we still do not know the lowest level of parasitemia that can remain stably in an individual The product should gain WHO and Gates Foundation support and yield LRE tests for other agents Eliminating Malaria from Haiti Reinventing DNA to Eradicate Endemic Parasites Firebird Biomolecular Sciences LLC University of Florida Steven A Benner John B Dame NARRATIVE With a population of less than million the absence of non human hosts and geographical isolation Haiti on the island of Hispaniola presents an ideal opportunity to eradicate Plasmodium falciparum the parasite that causes malaria The success of this program and such programs in other jurisdictions depends on the availability of an inexpensive assay to detect the parasite in human carriers individuals who are infected but display no symptoms This work will deliver such an assay one that targets falcimarum ribosomal RNA but is as simple to run as a rapid diagnostics test RDT This will benefit not only the Haitian population but also will help in the eradication of malaria elsewhere protect the US from malaria tourism and support work developing the science behind the epidemiology of human carrier reservoirs all while offering a paradigm for diagnostics in low resource environments