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Utilization of the rpoB Gene as a Specific Chromosomal Marker for Real-Time PCR Detection of Bacillus anthracis
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| Zeitschriftentitel: | Applied and Environmental Microbiology |
|---|---|
| Personen und Körperschaften: | , , , , , , |
| In: | Applied and Environmental Microbiology, 67, 2001, 8, S. 3720-3727 |
| Format: | E-Article |
| Sprache: | Englisch |
| veröffentlicht: |
American Society for Microbiology
|
| Schlagwörter: |
| author_facet |
Qi, Yuan Patra, Guy Liang, Xudong Williams, Leanne E. Rose, Sharon Redkar, Rajendra J. DelVecchio, Vito G. Qi, Yuan Patra, Guy Liang, Xudong Williams, Leanne E. Rose, Sharon Redkar, Rajendra J. DelVecchio, Vito G. |
|---|---|
| author |
Qi, Yuan Patra, Guy Liang, Xudong Williams, Leanne E. Rose, Sharon Redkar, Rajendra J. DelVecchio, Vito G. |
| spellingShingle |
Qi, Yuan Patra, Guy Liang, Xudong Williams, Leanne E. Rose, Sharon Redkar, Rajendra J. DelVecchio, Vito G. Applied and Environmental Microbiology Utilization of the rpoB Gene as a Specific Chromosomal Marker for Real-Time PCR Detection of Bacillus anthracis Ecology Applied Microbiology and Biotechnology Food Science Biotechnology |
| author_sort |
qi, yuan |
| spelling |
Qi, Yuan Patra, Guy Liang, Xudong Williams, Leanne E. Rose, Sharon Redkar, Rajendra J. DelVecchio, Vito G. 0099-2240 1098-5336 American Society for Microbiology Ecology Applied Microbiology and Biotechnology Food Science Biotechnology http://dx.doi.org/10.1128/aem.67.8.3720-3727.2001 <jats:title>ABSTRACT</jats:title> <jats:p> The potential use of <jats:italic>Bacillus anthracis</jats:italic> as a weapon of mass destruction poses a threat to humans, domesticated animals, and wildlife and necessitates the need for a rapid and highly specific detection assay. We have developed a real-time PCR-based assay for the specific detection of <jats:italic>B. anthracis</jats:italic> by taking advantage of the unique nucleotide sequence of the <jats:italic>B. anthracis rpoB</jats:italic> gene. Variable region 1 of the <jats:italic>rpoB</jats:italic> gene was sequenced from 36 <jats:italic>Bacillus</jats:italic> strains, including 16 <jats:italic>B. anthracis</jats:italic> strains and 20 other related bacilli, and four nucleotides specific for <jats:italic>B. anthracis</jats:italic> were identified. PCR primers were selected so that two <jats:italic>B. anthracis</jats:italic> -specific nucleotides were at their 3′ ends, whereas the remaining bases were specific to the probe region. This format permitted the PCR reactions to be performed on a LightCycler via fluorescence resonance energy transfer (FRET). The assay was found to be specific for 144 <jats:italic>B. anthracis</jats:italic> strains from different geographical locations and did not cross-react with other related bacilli (175 strains), with the exception of one strain. The PCR assay can be performed on isolated DNA as well as crude vegetative cell lysates in less than 1 h. Therefore, the <jats:italic>rpoB</jats:italic> -FRET assay could be used as a new chromosomal marker for rapid detection of <jats:italic>B. anthracis</jats:italic> . </jats:p> Utilization of the <i>rpoB</i> Gene as a Specific Chromosomal Marker for Real-Time PCR Detection of <i>Bacillus anthracis</i> Applied and Environmental Microbiology |
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10.1128/aem.67.8.3720-3727.2001 |
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American Society for Microbiology, 2001 |
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American Society for Microbiology, 2001 |
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| title |
Utilization of the rpoB Gene as a Specific Chromosomal Marker for Real-Time PCR Detection of Bacillus anthracis |
| title_unstemmed |
Utilization of the rpoB Gene as a Specific Chromosomal Marker for Real-Time PCR Detection of Bacillus anthracis |
| title_full |
Utilization of the rpoB Gene as a Specific Chromosomal Marker for Real-Time PCR Detection of Bacillus anthracis |
| title_fullStr |
Utilization of the rpoB Gene as a Specific Chromosomal Marker for Real-Time PCR Detection of Bacillus anthracis |
| title_full_unstemmed |
Utilization of the rpoB Gene as a Specific Chromosomal Marker for Real-Time PCR Detection of Bacillus anthracis |
| title_short |
Utilization of the rpoB Gene as a Specific Chromosomal Marker for Real-Time PCR Detection of Bacillus anthracis |
| title_sort |
utilization of the
<i>rpob</i>
gene as a specific chromosomal marker for real-time pcr detection of
<i>bacillus anthracis</i> |
| topic |
Ecology Applied Microbiology and Biotechnology Food Science Biotechnology |
| url |
http://dx.doi.org/10.1128/aem.67.8.3720-3727.2001 |
| publishDate |
2001 |
| physical |
3720-3727 |
| description |
<jats:title>ABSTRACT</jats:title>
<jats:p>
The potential use of
<jats:italic>Bacillus anthracis</jats:italic>
as a weapon of mass destruction poses a threat to humans, domesticated animals, and wildlife and necessitates the need for a rapid and highly specific detection assay. We have developed a real-time PCR-based assay for the specific detection of
<jats:italic>B. anthracis</jats:italic>
by taking advantage of the unique nucleotide sequence of the
<jats:italic>B. anthracis rpoB</jats:italic>
gene. Variable region 1 of the
<jats:italic>rpoB</jats:italic>
gene was sequenced from 36
<jats:italic>Bacillus</jats:italic>
strains, including 16
<jats:italic>B. anthracis</jats:italic>
strains and 20 other related bacilli, and four nucleotides specific for
<jats:italic>B. anthracis</jats:italic>
were identified. PCR primers were selected so that two
<jats:italic>B. anthracis</jats:italic>
-specific nucleotides were at their 3′ ends, whereas the remaining bases were specific to the probe region. This format permitted the PCR reactions to be performed on a LightCycler via fluorescence resonance energy transfer (FRET). The assay was found to be specific for 144
<jats:italic>B. anthracis</jats:italic>
strains from different geographical locations and did not cross-react with other related bacilli (175 strains), with the exception of one strain. The PCR assay can be performed on isolated DNA as well as crude vegetative cell lysates in less than 1 h. Therefore, the
<jats:italic>rpoB</jats:italic>
-FRET assay could be used as a new chromosomal marker for rapid detection of
<jats:italic>B. anthracis</jats:italic>
.
</jats:p> |
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| author | Qi, Yuan, Patra, Guy, Liang, Xudong, Williams, Leanne E., Rose, Sharon, Redkar, Rajendra J., DelVecchio, Vito G. |
| author_facet | Qi, Yuan, Patra, Guy, Liang, Xudong, Williams, Leanne E., Rose, Sharon, Redkar, Rajendra J., DelVecchio, Vito G., Qi, Yuan, Patra, Guy, Liang, Xudong, Williams, Leanne E., Rose, Sharon, Redkar, Rajendra J., DelVecchio, Vito G. |
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| description | <jats:title>ABSTRACT</jats:title> <jats:p> The potential use of <jats:italic>Bacillus anthracis</jats:italic> as a weapon of mass destruction poses a threat to humans, domesticated animals, and wildlife and necessitates the need for a rapid and highly specific detection assay. We have developed a real-time PCR-based assay for the specific detection of <jats:italic>B. anthracis</jats:italic> by taking advantage of the unique nucleotide sequence of the <jats:italic>B. anthracis rpoB</jats:italic> gene. Variable region 1 of the <jats:italic>rpoB</jats:italic> gene was sequenced from 36 <jats:italic>Bacillus</jats:italic> strains, including 16 <jats:italic>B. anthracis</jats:italic> strains and 20 other related bacilli, and four nucleotides specific for <jats:italic>B. anthracis</jats:italic> were identified. PCR primers were selected so that two <jats:italic>B. anthracis</jats:italic> -specific nucleotides were at their 3′ ends, whereas the remaining bases were specific to the probe region. This format permitted the PCR reactions to be performed on a LightCycler via fluorescence resonance energy transfer (FRET). The assay was found to be specific for 144 <jats:italic>B. anthracis</jats:italic> strains from different geographical locations and did not cross-react with other related bacilli (175 strains), with the exception of one strain. The PCR assay can be performed on isolated DNA as well as crude vegetative cell lysates in less than 1 h. Therefore, the <jats:italic>rpoB</jats:italic> -FRET assay could be used as a new chromosomal marker for rapid detection of <jats:italic>B. anthracis</jats:italic> . </jats:p> |
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| imprint | American Society for Microbiology, 2001 |
| imprint_str_mv | American Society for Microbiology, 2001 |
| institution | DE-Gla1, DE-Zi4, DE-15, DE-Rs1, DE-Pl11, DE-105, DE-14, DE-Ch1, DE-L229, DE-D275, DE-Bn3, DE-Brt1, DE-Zwi2, DE-D161 |
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| physical | 3720-3727 |
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| spelling | Qi, Yuan Patra, Guy Liang, Xudong Williams, Leanne E. Rose, Sharon Redkar, Rajendra J. DelVecchio, Vito G. 0099-2240 1098-5336 American Society for Microbiology Ecology Applied Microbiology and Biotechnology Food Science Biotechnology http://dx.doi.org/10.1128/aem.67.8.3720-3727.2001 <jats:title>ABSTRACT</jats:title> <jats:p> The potential use of <jats:italic>Bacillus anthracis</jats:italic> as a weapon of mass destruction poses a threat to humans, domesticated animals, and wildlife and necessitates the need for a rapid and highly specific detection assay. We have developed a real-time PCR-based assay for the specific detection of <jats:italic>B. anthracis</jats:italic> by taking advantage of the unique nucleotide sequence of the <jats:italic>B. anthracis rpoB</jats:italic> gene. Variable region 1 of the <jats:italic>rpoB</jats:italic> gene was sequenced from 36 <jats:italic>Bacillus</jats:italic> strains, including 16 <jats:italic>B. anthracis</jats:italic> strains and 20 other related bacilli, and four nucleotides specific for <jats:italic>B. anthracis</jats:italic> were identified. PCR primers were selected so that two <jats:italic>B. anthracis</jats:italic> -specific nucleotides were at their 3′ ends, whereas the remaining bases were specific to the probe region. This format permitted the PCR reactions to be performed on a LightCycler via fluorescence resonance energy transfer (FRET). The assay was found to be specific for 144 <jats:italic>B. anthracis</jats:italic> strains from different geographical locations and did not cross-react with other related bacilli (175 strains), with the exception of one strain. The PCR assay can be performed on isolated DNA as well as crude vegetative cell lysates in less than 1 h. Therefore, the <jats:italic>rpoB</jats:italic> -FRET assay could be used as a new chromosomal marker for rapid detection of <jats:italic>B. anthracis</jats:italic> . </jats:p> Utilization of the <i>rpoB</i> Gene as a Specific Chromosomal Marker for Real-Time PCR Detection of <i>Bacillus anthracis</i> Applied and Environmental Microbiology |
| spellingShingle | Qi, Yuan, Patra, Guy, Liang, Xudong, Williams, Leanne E., Rose, Sharon, Redkar, Rajendra J., DelVecchio, Vito G., Applied and Environmental Microbiology, Utilization of the rpoB Gene as a Specific Chromosomal Marker for Real-Time PCR Detection of Bacillus anthracis, Ecology, Applied Microbiology and Biotechnology, Food Science, Biotechnology |
| title | Utilization of the rpoB Gene as a Specific Chromosomal Marker for Real-Time PCR Detection of Bacillus anthracis |
| title_full | Utilization of the rpoB Gene as a Specific Chromosomal Marker for Real-Time PCR Detection of Bacillus anthracis |
| title_fullStr | Utilization of the rpoB Gene as a Specific Chromosomal Marker for Real-Time PCR Detection of Bacillus anthracis |
| title_full_unstemmed | Utilization of the rpoB Gene as a Specific Chromosomal Marker for Real-Time PCR Detection of Bacillus anthracis |
| title_short | Utilization of the rpoB Gene as a Specific Chromosomal Marker for Real-Time PCR Detection of Bacillus anthracis |
| title_sort | utilization of the <i>rpob</i> gene as a specific chromosomal marker for real-time pcr detection of <i>bacillus anthracis</i> |
| title_unstemmed | Utilization of the rpoB Gene as a Specific Chromosomal Marker for Real-Time PCR Detection of Bacillus anthracis |
| topic | Ecology, Applied Microbiology and Biotechnology, Food Science, Biotechnology |
| url | http://dx.doi.org/10.1128/aem.67.8.3720-3727.2001 |