Abstract
The persistence of DNA after the cell death causes a major issue in aspects of medical or biological studies. The signal from viable bacterial cells cannot be distinguished from the dead cells in the conventional DNA-based detection methods. In the present study, the loop-mediated isothermal amplification (LAMP) method combined with the ethidium monoazide (EMA) treatment was applied for specific detection of viable, but not dead, Salmonella cells. For this method (EMA-LAMP), we designed a series of primers, which recognize six distinct sequences of the target invA gene conserved in Salmonella. The invA gene of the viable cells was remarkably amplified within 1 hr when as small amounts as 100 fg of DNA was subjected to EMA-LAMP. Because EMA selectively penetrated into the dead cells and bound covalently to DNA, the gene of the dead cells could not be amplified. This study offers a novel DNA-based method to distinguish the viable bacterial cells from the dead cells.
| Original language | English |
|---|---|
| Pages (from-to) | 820-824 |
| Number of pages | 5 |
| Journal | Journal of Health Science |
| Volume | 55 |
| Issue number | 5 |
| DOIs | |
| Publication status | Published - Oct 2009 |
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Keywords
- Ethidium monoazide
- Loop-mediated isothermal amplification
- Salmonella
- Viable cell
ASJC Scopus subject areas
- Toxicology
- Health, Toxicology and Mutagenesis
Cite this
Specific detection of viable Salmonella cells by an ethidium monoazide-loop mediated isothermal amplification (EMA-LAMP) method. / Lu, Yuxia; Yang, Weiqing; Shi, Lei; Li, N.; Alam, Muhammad Jahangir; Guo, Siyuan; Miyoshi, Shin-ichi.
In: Journal of Health Science, Vol. 55, No. 5, 10.2009, p. 820-824.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Specific detection of viable Salmonella cells by an ethidium monoazide-loop mediated isothermal amplification (EMA-LAMP) method
AU - Lu, Yuxia
AU - Yang, Weiqing
AU - Shi, Lei
AU - Li, N.
AU - Alam, Muhammad Jahangir
AU - Guo, Siyuan
AU - Miyoshi, Shin-ichi
PY - 2009/10
Y1 - 2009/10
N2 - The persistence of DNA after the cell death causes a major issue in aspects of medical or biological studies. The signal from viable bacterial cells cannot be distinguished from the dead cells in the conventional DNA-based detection methods. In the present study, the loop-mediated isothermal amplification (LAMP) method combined with the ethidium monoazide (EMA) treatment was applied for specific detection of viable, but not dead, Salmonella cells. For this method (EMA-LAMP), we designed a series of primers, which recognize six distinct sequences of the target invA gene conserved in Salmonella. The invA gene of the viable cells was remarkably amplified within 1 hr when as small amounts as 100 fg of DNA was subjected to EMA-LAMP. Because EMA selectively penetrated into the dead cells and bound covalently to DNA, the gene of the dead cells could not be amplified. This study offers a novel DNA-based method to distinguish the viable bacterial cells from the dead cells.
AB - The persistence of DNA after the cell death causes a major issue in aspects of medical or biological studies. The signal from viable bacterial cells cannot be distinguished from the dead cells in the conventional DNA-based detection methods. In the present study, the loop-mediated isothermal amplification (LAMP) method combined with the ethidium monoazide (EMA) treatment was applied for specific detection of viable, but not dead, Salmonella cells. For this method (EMA-LAMP), we designed a series of primers, which recognize six distinct sequences of the target invA gene conserved in Salmonella. The invA gene of the viable cells was remarkably amplified within 1 hr when as small amounts as 100 fg of DNA was subjected to EMA-LAMP. Because EMA selectively penetrated into the dead cells and bound covalently to DNA, the gene of the dead cells could not be amplified. This study offers a novel DNA-based method to distinguish the viable bacterial cells from the dead cells.
KW - Ethidium monoazide
KW - Loop-mediated isothermal amplification
KW - Salmonella
KW - Viable cell
UR - http://www.scopus.com/inward/record.url?scp=70350236729&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=70350236729&partnerID=8YFLogxK
U2 - 10.1248/jhs.55.820
DO - 10.1248/jhs.55.820
M3 - Article
AN - SCOPUS:70350236729
VL - 55
SP - 820
EP - 824
JO - Journal of Health Science
JF - Journal of Health Science
SN - 1344-9702
IS - 5
ER -