Two-Component Direct Fluorescent-Antibody Assay for Rapid Identification of Bacillus anthracis
نویسندگان
چکیده
A two-component direct fluorescent-antibody (DFA) assay, using fluorescein-labeled monoclonal antibodies specific to the Bacillus anthracis cell wall (CW-DFA) and capsule (CAP-DFA) antigens, was evaluated and validated for rapid identification of B. anthracis. We analyzed 230 B. anthracis isolates; 228 and 229 were positive by CW-DFA and CAP-DFA assays, respectively. We also tested 56 non-B. anthracis strains; 10 B. cereus and 2 B. thuringiensis were positive by the CW-DFA assay, and 1 B. megaterium strain was positive by CAP-DFA. Analysis of the combined DFA results identified 227 of 230 B. anthracis isolates; all 56 strains of the other Bacillus spp. were negative. Both DFA assays tested positive on 14 of 26 aging clinical specimens from the 2001 anthrax outbreak investigation. The two-component DFA assay is a sensitive, specific, and rapid confirmatory test for B. anthracis in cultures and may be useful directly on clinical specimens.
منابع مشابه
Identification and Characterization of Bacillus anthracis Spores by Flow Cytometry
Rapid and accurate detection of Bacillus anthracis, the causative agent of anthrax, remains an active area of research due to the continued threat of bioterrorist attack. The ability to differentiate Bacillus anthracis spores from spores belonging to other Bacillus species is important for the development of spore-based detection methods. Furthermore, not all Bacillus anthracis strains are full...
متن کاملDevelopment of antibodies against anthrose tetrasaccharide for specific detection of Bacillus anthracis spores.
Methods for the immunological detection of Bacillus anthracis in various environmental samples and the discrimination of B. anthracis from other members of the B. cereus group are not yet well established. To generate specific discriminating antibodies, we immunized rabbits, mice, and chickens with inactivated B. anthracis spores and, additionally, immunized rabbits and mice with the tetrasacch...
متن کاملDetection of Bacillus anthracis DNA by LightCycler PCR.
Anthrax is a zoonotic disease that is also well recognized as a potential agent of bioterrorism. Routine culture and biochemical testing methods are useful for the identification of Bacillus anthracis, but a definitive identification may take 24 to 48 h or longer and may require that specimens be referred to another laboratory. Virulent isolates of B. anthracis contain two plasmids (pX01 and pX...
متن کاملReal-time PCR assay for a unique chromosomal sequence of Bacillus anthracis.
Real-time PCR has become an important method for the rapid identification of Bacillus anthracis since the 2001 anthrax mailings. Most real-time PCR assays for B. anthracis have been developed to detect virulence genes located on the pXO1 and pXO2 plasmids. In contrast, only two published chromosomal targets exist, the rpoB gene and the gyrA gene. In the present study, subtraction-hybridization ...
متن کاملDevelopment and implementation of a single-chain Fv antibody for specific detection of Bacillus anthracis spores.
A single-chain Fv (scFv) antibody was developed and applied for efficient and specific detection of Bacillus anthracis spores. The antibody was isolated from a phage display library prepared from spleens of mice immunized with a water-soluble extract of the outer membrane of the B. anthracis spore (exosporium). The library (7 x 10(6) PFU) was biopanned against live, native B. anthracis ATCC del...
متن کامل