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Comparison of two suspension arrays for simultaneous detection of five biothreat bacterial in powder samples.

Yang Y, Wang J, Wen H, Liu H - J. Biomed. Biotechnol. (2012)

Bottom Line: Universal primers were used to amplify highly conserved region located within the 16S rRNA amplicon, followed by hybridized to pathogen-specific probes for identification of these five organisms.Both of the two arrays are validated to be flexible and sensitive for simultaneous detection of bioterrorism bacteria.The two suspension arrays can be utilized to detect Bacillus anthracis sterne spore and Yersinia pestis EV76 from mimic "write powder" samples, they also proved that the suspension array system will be valuable tools for diagnosis of bacterial biothreat agents in environmental samples.

View Article: PubMed Central - PubMed

Affiliation: Institute of Health Quarantine, Chinese Academy of Inspection and Quarantine, Beijing 100123, China.

ABSTRACT
We have developed novel Bio-Plex assays for simultaneous detection of Bacillus anthracis, Yersinia pestis, Brucella spp., Francisella tularensis, and Burkholderia pseudomallei. Universal primers were used to amplify highly conserved region located within the 16S rRNA amplicon, followed by hybridized to pathogen-specific probes for identification of these five organisms. The other assay is based on multiplex PCR to simultaneously amplify five species-specific pathogen identification-targeted regions unique to individual pathogen. Both of the two arrays are validated to be flexible and sensitive for simultaneous detection of bioterrorism bacteria. However, universal primer PCR-based array could not identify Bacillus anthracis, Yersinia pestis, and Brucella spp. at the species level because of the high conservation of 16S rDNA of the same genus. The two suspension arrays can be utilized to detect Bacillus anthracis sterne spore and Yersinia pestis EV76 from mimic "write powder" samples, they also proved that the suspension array system will be valuable tools for diagnosis of bacterial biothreat agents in environmental samples.

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Detectability for white powder mixture. (a) Bacillus anthracis sterne spore 102 ~ 109 cells/0.1 g detected in white powder. (b) Yersinia pestis Ev76 5 × 101 ~ 5 × 108 cfu/0.1 g detected in white powder.
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Related In: Results  -  Collection


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fig5: Detectability for white powder mixture. (a) Bacillus anthracis sterne spore 102 ~ 109 cells/0.1 g detected in white powder. (b) Yersinia pestis Ev76 5 × 101 ~ 5 × 108 cfu/0.1 g detected in white powder.

Mentions: As shown in Figure 5, we identified B. anthracis spore, and Yersinia pestis EV76 in simulated “white powder”, 10000 cells/0.1 g powder of B. anthracis spore and 500 cfu/0.1 g powder of Yersinia pestis EV76 showed positive signals when tested by both two assays, which are significantly below the median lethal dose (LD50) of 8000~10000 cfu Bacillus anthracis [10] and 3000 cfu Yersinia pestis [11]. The results suggested these two types of arrays have enough detective ability to detect suspect bioterrorism agents from white powders with high sensitivity and good dynamic detection range.


Comparison of two suspension arrays for simultaneous detection of five biothreat bacterial in powder samples.

Yang Y, Wang J, Wen H, Liu H - J. Biomed. Biotechnol. (2012)

Detectability for white powder mixture. (a) Bacillus anthracis sterne spore 102 ~ 109 cells/0.1 g detected in white powder. (b) Yersinia pestis Ev76 5 × 101 ~ 5 × 108 cfu/0.1 g detected in white powder.
© Copyright Policy - open-access
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3368695&req=5

fig5: Detectability for white powder mixture. (a) Bacillus anthracis sterne spore 102 ~ 109 cells/0.1 g detected in white powder. (b) Yersinia pestis Ev76 5 × 101 ~ 5 × 108 cfu/0.1 g detected in white powder.
Mentions: As shown in Figure 5, we identified B. anthracis spore, and Yersinia pestis EV76 in simulated “white powder”, 10000 cells/0.1 g powder of B. anthracis spore and 500 cfu/0.1 g powder of Yersinia pestis EV76 showed positive signals when tested by both two assays, which are significantly below the median lethal dose (LD50) of 8000~10000 cfu Bacillus anthracis [10] and 3000 cfu Yersinia pestis [11]. The results suggested these two types of arrays have enough detective ability to detect suspect bioterrorism agents from white powders with high sensitivity and good dynamic detection range.

Bottom Line: Universal primers were used to amplify highly conserved region located within the 16S rRNA amplicon, followed by hybridized to pathogen-specific probes for identification of these five organisms.Both of the two arrays are validated to be flexible and sensitive for simultaneous detection of bioterrorism bacteria.The two suspension arrays can be utilized to detect Bacillus anthracis sterne spore and Yersinia pestis EV76 from mimic "write powder" samples, they also proved that the suspension array system will be valuable tools for diagnosis of bacterial biothreat agents in environmental samples.

View Article: PubMed Central - PubMed

Affiliation: Institute of Health Quarantine, Chinese Academy of Inspection and Quarantine, Beijing 100123, China.

ABSTRACT
We have developed novel Bio-Plex assays for simultaneous detection of Bacillus anthracis, Yersinia pestis, Brucella spp., Francisella tularensis, and Burkholderia pseudomallei. Universal primers were used to amplify highly conserved region located within the 16S rRNA amplicon, followed by hybridized to pathogen-specific probes for identification of these five organisms. The other assay is based on multiplex PCR to simultaneously amplify five species-specific pathogen identification-targeted regions unique to individual pathogen. Both of the two arrays are validated to be flexible and sensitive for simultaneous detection of bioterrorism bacteria. However, universal primer PCR-based array could not identify Bacillus anthracis, Yersinia pestis, and Brucella spp. at the species level because of the high conservation of 16S rDNA of the same genus. The two suspension arrays can be utilized to detect Bacillus anthracis sterne spore and Yersinia pestis EV76 from mimic "write powder" samples, they also proved that the suspension array system will be valuable tools for diagnosis of bacterial biothreat agents in environmental samples.

Show MeSH
Related in: MedlinePlus