Abstract
Inhalational anthrax is a serious biothreat. Effective antibiotic treatment of inhalational anthrax requires early diagnosis; the further the disease has progressed, the less the likelihood for cure. Current means for diagnosis such as blood culture require several days to a result and require advanced laboratory infrastructure. An alternative approach to diagnosis is detection of a Bacillus anthracis antigen that is shed into blood and can be detected by rapid immunoassay. The goal of the study was to evaluate detection of poly-γ-D-glutamic acid (PGA), the capsular antigen of B. anthracis, as a biomarker surrogate for blood culture in a rabbit model of inhalational anthrax. The mean time to a positive blood culture was 26 ± 5.7 h (mean ± standard deviation), whereas the mean time to a positive ELISA was 22 ± 4.2 h; P = 0.005 in comparison with blood culture. A lateral flow immunoassay was constructed for detection of PGA in plasma at concentrations of less than 1 ng PGA/ml. Use of the lateral flow immunoassay for detection of PGA in the rabbit model found that antigen was detected somewhat earlier than the earliest time point at which the blood culture became positive. The low cost, ease of use, and rapid time to result of the lateral flow immunoassay format make an immunoassay for PGA a viable surrogate for blood culture for detection of infection in individuals who have a likelihood of exposure to B. anthracis.
Highlights
Bacillus anthracis, the causative agent of inhalational anthrax, is a major biothreat
The murine IgG3 monoclonal antibodies (mAbs) F26G3 was produced from BALB/c mice that were immunized with B. licheniformis poly-γ-D-glutamic acid (PGA) in combination with CD40 agonist antibodies [3,9,10]. mAb 8B10 was produced by subcutaneous immunization of CD1 mice with a synthetic γDPGA oligomer coupled to KLH and emulsified with TiterMax Gold (Titermax, Norcross, GA)
An enzyme-linked immunosorbent assay (ELISA) was constructed with the IgG3 anti-PGA mAb F26G3 and was used to quantify PGA levels in rabbit plasma and urine collected in a time-point series following inhalational exposure to B. anthracis Ames spores
Summary
The causative agent of inhalational anthrax, is a major biothreat. Lucey has proposed a four-stage classification for the progression of inhalational anthrax: asymptomatic, early-prodromal, intermediate-progressive and late-fulminant [1]. The efficacy for treatment of inhalational anthrax depends on the stage at which antibiotics are administered. The further the disease has progressed, the less the likelihood for cure. Few patients will survive if untreated disease progresses to the late-fulminant stage.
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