Test and Evaluation of Field-Deployable Infectious Disease Diagnostic Assays in Support of the Joint Biological Agent Identification and Diagnosis System (JBAIDS): Malaria (Plasmodium/JBAIDS)
Abstract
A Plasmodium genus dual-fluorogenic, hydrolysis probe (TaqMan) polymerase chain reaction (PCR) assay was developed using real-time PCR instrumentation (MiniOpticon Real-Time PCR System, Bio-Rad, Hercules, CA). The Plasmodium genus assay is sensitive, specific, and provides rapid detection from infected mosquitoes and malaria patient blood. Operational applications are FHP disease surveillance and ultimately molecular-based diagnostics. A unique ISS rRNA gene sequence conserved in Plasmodium species was targeted for the design of oligonucleotide primers and probe. The LoD of the Plasmodium genus TaqMan assay was established at 10 genome equivalents using P. vivax plasmid and P. falciparum plasmid. The assay was 100% (17/17) concordant in testing using a diverse panel of Plasmodium species and strains prepared as plasmid control DNA. The assay was 100% (5/5) sensitive and 100% (13/13) specific in testing with a diverse panel of human malaria Plasmodium species and non-malaria pathogens. No cross-reaction occurred with human blood samples. Transfer to the RAPID/JBAIDS presents no technical challenges.
Document Details
- Document Type
- Technical Report
- Publication Date
- May 31, 2012
- Accession Number
- ADA562427
Entities
People
- James A. Swaby
- James C. Mcavin
- Ratree Takhampunya
Organizations
- 59th Medical Wing