Recent literature review of soil processing methods for recovery of Bacillus anthracis spores

The detection and quantification of Bacillus thuringiensis spores from soil and swabs using quantitative PCR as a model system for routine diagnostics of Bacillus anthracis

“…; Silvestri et al . ). Isolation of bacillary spores from sand and sandy soil is mostly more effective compared to other soil types (Agarwal et al .…”

“…In soil samples, direct processing is complicated by the presence of other spore‐forming bacterial species and by soil compounds, such as humic acids, which are known to be PCR inhibitors (Silvestri et al . ). Although the use of the direct method can yield good quality DNA, the total amount of isolated DNA can be quite low using this technique.…”

“…Direct processing is based on isolation of spores directly from environmental samples without the necessity for prior pre-extraction or concentration of the sample. In soil samples, direct processing is complicated by the presence of other sporeforming bacterial species and by soil compounds, such as humic acids, which are known to be PCR inhibitors (Silvestri et al 2015). Although the use of the direct method can yield good quality DNA, the total amount of isolated DNA can be quite low using this technique.…”

Rapid Detection of Viable Bacillus anthracis Spores in Environmental Samples by Using Engineered Reporter Phages

“…With the advent of more rapid and less expensive wholegenome sequencing approaches, discrimination between closely related species and strains is readily attainable (16,17). Nevertheless, chemical constituents of soil (such as organics, humic acids, and/or heavy metals) often interfere with nucleic acid-based chemistry and make direct detection of B. anthracis extremely difficult (18)(19)(20)(21). Consequently, the majority of detection assays incorporate sample processing methods to separate, concentrate, and purify B. anthracis spores from soil prior to DNA extraction (21).…”

“…Nevertheless, chemical constituents of soil (such as organics, humic acids, and/or heavy metals) often interfere with nucleic acid-based chemistry and make direct detection of B. anthracis extremely difficult (18)(19)(20)(21). Consequently, the majority of detection assays incorporate sample processing methods to separate, concentrate, and purify B. anthracis spores from soil prior to DNA extraction (21). Further complications arise during clearance monitoring as small numbers of viable spores must be recognized amid a landscape of nonviable spores.…”