Bacillus spore detection

S. Farquharson, A. Gift, P. Maksymiuk and F. Inscore, Rapid dipicolinic acid extraction Ifom Bacillus spores detected by surface-enhanced Raman spectroscopy, Appl. Spectrosc. 58, 351... 

Goodacre, R. Shann, B. Gilbert, R. J. Timmis, E. M. McGovern, A. C. Alsberg, B. K. Kell, D. B. Logan, N. A. Detection of the dipicolinic acid biomarker in Bacillus spores using Curie-point pyrolysis mass spectrometry and fourier transform infrared spectroscopy Anal. Chem. 2000,72,119-127. 

Snyder, A. P Maswadeh, W. M. Parsons, J. A. Tripathi, A. Meuzelaar, H. L. C. Dworzanski, J. P. Kim, M. G. Field detection of Bacillus spore aerosols with standalone pyrolysis-gas chromatography-ion mobility spectrometry. Field Anal. Chem. Technol. 1999, 3, 315-326. 

Tumbough C, Kearney J. 1998. Capture and Detection of Bacillus Spores. Presentation at the Defense Advanced Research Projects Agency Meeting on Bio-surveillance Providing Detection in the New Millenium. Johns Hopkins University Applied Physics Laboratory, Laurel, MD, February 11. 

CaDPA) has a high affinity for lanthanide ions and in particular terbium(III) complexes are considered the best probes for the detection of this species which is a unique biomarker for bacillus spores such as B. anthracis, known as a potential agent for biological terroristic attacks. 

In what follows, the uses of MS for pathogen detection would be demonstrated in the examples of intact Bacillus spore species, as well as Plasmodium parasites— the causative agent of malaria. The types of characteristic biomarker molecules detected by MS for these two very different cases would be discussed as well as the factors influencing mass spectra of intact organisms. 

Anthrax is an infection disease caused by Bacillus anthracis that requires medical attention within 24-48 hours after initial inhalation of more than 10 B, anthracis spores (/). Therefore, the rapid detection of B, anthracis spores in the environment prior to infection is an extremely important goal for human safety. Structurally, a spore consists of a central core cell surrounded by various protective layers. CaDPA is located in these protective layers and accounts for -10% of the spore s dry weight (2) making it is a useful biomarker for bacillus spores (5). 

Elhanany, E. Barak, R. Fisher, M. Kobiler, D. Altboum, Z. Detection of specific Bacillus anthracis spore biomakers by matrix-assisted desorption/ionization time-of-flight mass spectrometry. Rapid Comm. Mass Spectrom. 2001,15, 2110-2116. 

Events involving deliberate or accidental distribution of bacterial pathogens into our everyday environment have clearly defined the need for a sensitive, specific, and rapid method of bacterial detection. Bioterrorism was first introduced in the United States in 1984 with the Salmonella typhimurium attack in The Dalles, Oregon, by a cult group attempting to affect a local election.1 As a result of this act 751 people contracted salmonellosis, which totally overwhelmed the hospitals and medical clinics with patients. Later our society became keenly aware of the potential of bioterrorism during the last four months of 2001 when Bacillus anthracis (anthrax) spores were sent through the US mail in an envelope to several locations. These events had... 

Bruno J.G., Yu H., Immunomagnetic electrochemiluminescent detection of Bacillus anthracis spores in soil matrices, Appl. Environ. Micro. 1996 62 3474-3476. 

Stratis-Cullum D.N., Griffin G.D., Mobley J., Vass A.A., Vo-Dinh T., A miniature biochip system for detection of aerosolized Bacillus globigii spores, Anal. Chem. 2003 75 275-280. 

E.T. Arakawa, N.V. Lavrik and P.G. Datskos, Detection of anthrax stimulants with microcalorimetric spectroscopy Bacillus subtilis and Bacillus cereus spores, Appl. Opt., 42 (2003) 1757-1762. 

Petrov, G. 1., Yakovlev, V. V., Sokolov, A., and Scully, M. O. 2005. Detection of Bacillus subti-lis spores in water by means of broadband coherent anti-Stokes Raman spectroscopy. Opt. Express 13 9537 2. 

The marker substance for Bacillus endospores is calcium dipicolinate (CaDPA). One approach is the isolation of CaDPA from a minimal amount of spores and its subsequent identification by SERS spectroscopy [97]. The detection of CaDPA in endospores was used for the detection of minimal amount of spores [98] even in automated mail sorting systems [99]. The detection of CaDPA does not give any information about the Bacillus species present. The investigation of the components in the outmost layers of the endospores, the outer sporecoat, or the exosporium will lead to more precise information,... 

Real-time PCR was also performed by on-chip thermal cycling for the detection of Hantavirus, HIV, orthopoxviruses (266-281 bp), Borrelia burgdorferi, human (3-actin (294 bp), and the human complement C6 gene (73 bp) [944]. In another report, real-time PCR was performed for DNA extracted from Bacillus subtilis in a plastic microfluidic cassette. The spore samples (lOVmL) were sampled, filtered, and sonicated in the presence of 6-pm glass beads for spore disruption to release the DNA [945],... 

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