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Discrimination of Spore-Forming Bacilli Using spoIVAA method of discriminating between spore-forming and non-spore-forming bacteria is based on a combination of simultaneous sporulation-specific and non-sporulation-specific quantitative polymerase chain reactions (Q-PCRs). The method was invented partly in response to the observation that for the purposes of preventing or reducing biological contamination affecting many human endeavors, ultimately, only the spore-forming portions of bacterial populations are the ones that are problematic (or, at least, more problematic than are the non-spore-forming portions). In some environments, spore-forming bacteria constitute small fractions of the total bacterial populations. The use of sporulation-specific primers in Q-PCR affords the ability to assess the spore-forming fraction of a bacterial population present in an environment of interest. This assessment can provide a more thorough and accurate understanding of the bacterial contamination in the environment, thereby making it possible to focus contamination- testing, contamination-prevention, sterilization, and decontamination resources more economically and efficiently. The method includes the use of sporulation-specific primers in the form of designed, optimized deoxyribonucleic acid (DNA) oligonucleotides specific for the bacterial spoIVA gene (see table). [In "spoIVA," "IV" signifies Roman numeral four and the entire quoted name refers to gene A for the fourth stage of sporulation.] These primers are mixed into a PCR cocktail with a given sample of bacterial cells. A control PCR cocktail into which are mixed universal 16S rRNA primers is also prepared. ["16S rRNA" denotes a ribosomal ribonucleic acid (rRNA) sequence that is common to all organisms.] Following several cycles of heating and cooling according to the PCR protocol to amplify amounts of DNA molecules, the amplification products can be analyzed to determine the types of bacterial cells present within the samples. If the amplification product is strong, relative to the product of a control PCR sequence, then it is concluded that the bacterial population in the sample consists predominantly of spore-forming cells. If the amplification product is weak or nonexistent, then it is concluded that the bacterial population in the sample consists predominantly or entirely of non-spore-forming cells.
Document ID
20090011190
Acquisition Source
Jet Propulsion Laboratory
Document Type
Other - NASA Tech Brief
External Source(s)
http://www.techbriefs.com/component/content/article/5010
Authors
Venkateswaran, Kasthuri (California Inst. of Tech. Pasadena, CA, United States)
LaDuc, Myron (California Inst. of Tech. Pasadena, CA, United States)
Stuecker, Tara (California Inst. of Tech. Pasadena, CA, United States)
Date Acquired
August 24, 2013
Publication Date
March 1, 2009
Publication Information
Publication: NASA Tech Briefs, March 2009
Subject Category
Man/System Technology And Life Support
Report/Patent Number
NPO-44296
Distribution Limits
Public
Copyright
Public Use Permitted.

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IDRelationTitle20090011183Collected WorksNASA Tech Briefs, March 2009
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