Detection of Bacillus Spores Using PCR and FTA Filters

Lampel, Keith A.; Dyer, Deanne; Kornegay, Leroy; Orlandi, Palmer A.

doi:10.4315/0362-028x-67.5.1036

Cited by 18 publications

(6 citation statements)

References 9 publications

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“…A number of serological and molecular techniques exist for the detection of B. cereus or other members of the genus Bacillus (Chen et al 2001;Hansen and Hendriksen 2001;Uyttendaele and Debevere 2003;Lampel et al 2004;Kim et al 2005;Varughese et al 2007). These nongrowth-based methods are rapid and provide an unequivocal identification of the organism without the need to perform further tests, as is the case with plate counting (Mathews 2003;Zwirglmaier 2005).…”

Section: Detectionmentioning

confidence: 99%

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The potential of flow cytometry in the study of Bacillus cereus

Cronin

Wilkinson

2010

Journal of Applied Microbiology

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Summary Flow cytometry (FCM) is a rapid method allowing the acquisition of multiparametric data from thousands of individual cells within a sample. As well as measuring the intrinsic light scattering properties of cells, a plethora of fluorescent dyes may be employed to yield information on macromolecule content, surface antigens present or physiological status. Despite FCM’s indispensability within other fields e.g. immunology, it is underutilized within microbiological research. In this review, a strong case is presented for the potential of FCM in the study of Gram‐positive spore‐former, Bacillus cereus. Previous reports where FCM was successfully used in the study of B. cereus are reviewed along with relevant studies involving other members of the genus. Under headings reflecting common research themes associated with B. cereus, specific instances where FCM has generated novel data, providing a unique insight into the organism, are discussed. Further applications are posited, based on the authors’ own research with FCM and B. cereus and work extant in the broader field of microbial cytometry. The authors conclude that, while the expense of equipment and reagents is an undeniable disadvantage, FCM is a technique capable of generating significantly novel data and allows the design and execution of experiments that are not possible with any other technique.

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Section: Detectionmentioning

confidence: 99%

“…A number of serological and molecular techniques exist for the detection of B. cereus or other members of the genus Bacillus (Chen et al. 2001; Hansen and Hendriksen 2001; Uyttendaele and Debevere 2003; Lampel et al. 2004; Kim et al.…”

Section: Common Themes In Bacillus Cereus Research and The Utility Ofmentioning

confidence: 99%

The potential of flow cytometry in the study of Bacillus cereus

Cronin

Wilkinson

2010

Journal of Applied Microbiology

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“…FTA filters have been used for the detection of Bacillus subtilis, Bacillus cereus, and Bacillus megaterium spores using nested PCR. The reported sensitivity of this method is 53 spores in the first round of PCR, and 5 spores after the second nested PCR round (169). The preparation of template DNA from biothreat agents such as Bacillus anthracis spores using FTA filters should be similar although samples such as powder have not yet been tested.…”

Section: Methods For Sample Processingmentioning

confidence: 99%

Current and Developing Technologies for Monitoring Agents of Bioterrorism and Biowarfare

et al. 2005

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SUMMARY Recent events have made public health officials acutely aware of the importance of rapidly and accurately detecting acts of bioterrorism. Because bioterrorism is difficult to predict or prevent, reliable platforms to rapidly detect and identify biothreat agents are important to minimize the spread of these agents and to protect the public health. These platforms must not only be sensitive and specific, but must also be able to accurately detect a variety of pathogens, including modified or previously uncharacterized agents, directly from complex sample matrices. Various commercial tests utilizing biochemical, immunological, nucleic acid, and bioluminescence procedures are currently available to identify biological threat agents. Newer tests have also been developed to identify such agents using aptamers, biochips, evanescent wave biosensors, cantilevers, living cells, and other innovative technologies. This review describes these current and developing technologies and considers challenges to rapid, accurate detection of biothreat agents. Although there is no ideal platform, many of these technologies have proved invaluable for the detection and identification of biothreat agents.

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“…Specifically, they have been used in template preparation for PCR detection of pathogenic microorganisms. For example, Lampel et al (2000), Orlandi and Lampel (2000), Lampel and Orlandi (2002), Hide et al (2003), Warren (2003), Lampel et al (2004) and Warren et al (2005) used PCR to detect pathogenic microorganisms from different foods using FTA cards. FTA technique is a simple, rapid, precise and highly sensitive even to low count of bacterial cells in samples.…”

Section: Introductionmentioning

confidence: 99%

The Sensitivity of PCR to the Number of Lactococcus garvieae in Different Concentrations of NaCl: FTA Technique

Alrabadi¹

2012

IJB

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The major objective of this study is to investigate the sensitivity of PCR to the number of Lactococcus garvieae (L. garvieae) in sterilized milk containing different concentrations of NaCl using FTA technique for genomic DNA extraction. This technique is a simple, rapid, precise and highly sensitive even to low count of bacterial cells in samples. The study used M17 medium to determine the number of bacterial cells, thereafter, FTA method was used for genomic DNA extraction. The results using M17 medium, indicated that the L. garvieae increased in all concentrations of NaCl during an incubation period of 24 hours, on the other hand, pH value decreased. However, the increase in bacterial count was statistically significant only on 0, 2% and 4% of NaCl concentrations. In conjunction with this, the PCR showed sensitivity to both low and high numbers of bacterial cells in the samples. Increasing bacterial numbers led to an increase in the amount of DNA extracted via FTA cards and therefore, the intensity and clarity of bands of PCR reaction was also increased. This is the first study that uses FTA technique for DNA extraction in dairy products. Using FTA cards and subjecting to PCR in quantifying the bacterial growth is a very important issue because of its direct application in the field of milk, agricultural and animal products research and development.

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Detection of Bacillus Spores Using PCR and FTA Filters

Cited by 18 publications

References 9 publications

The potential of flow cytometry in the study of Bacillus cereus

The potential of flow cytometry in the study of Bacillus cereus

Current and Developing Technologies for Monitoring Agents of Bioterrorism and Biowarfare

The Sensitivity of PCR to the Number of Lactococcus garvieae in Different Concentrations of NaCl: FTA Technique

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