Abstract
Brucella spp. are facultative intracellular bacteria that cause zoonotic disease of brucellosis worldwide. Livestock that are most vulnerable to brucellosis include cattle, goats, and pigs. Brucella spp. cause serious health problems to humans and animals and economic losses to the livestock industry. Traditional methods for detection of Brucella spp. take 48-72 h (Kumar et al., J Commun Dis 29:131-137, 1997; Barrouin-Melo et al., Res Vet Sci 83:340-346, 2007) that do not meet the food industry's need of rapid detection. Therefore, there is an urgent need of fast, specific, sensitive, and inexpensive method for diagnosing of Brucella spp. Loop-mediated isothermal amplification (LAMP) is a method to amplify nucleic acid at constant temperatures. Amplification can be detected by visual detection, fluorescent stain, turbidity, and electrophoresis. We targeted at the Brucella-specific gene omp25 and designed LAMP primers for detection of Brucella spp. Amplification of DNA with Bst DNA polymerase can be completed at 65 C in 60 min. Amplified products can be detected by SYBR Green I stain and 2.0 % agarose gel electrophoresis. The LAMP method is feasible for detection of Brucella spp. from blood and milk samples.
| Original language | English (US) |
|---|---|
| Title of host publication | Methods in Molecular Biology |
| Pages | 99-108 |
| Number of pages | 10 |
| Volume | 1039 |
| DOIs | |
| State | Published - 2013 |
Publication series
| Name | Methods in Molecular Biology |
|---|---|
| Volume | 1039 |
| ISSN (Print) | 10643745 |
Fingerprint
Keywords
- Blood
- Brucella
- Detection
- LAMP
- Loop-mediated isothermal amplification
- Milk
ASJC Scopus subject areas
- Molecular Biology
- Genetics
Cite this
Rapid detection of brucella spp. using loop-mediated isothermal amplification (LAMP). / Chen, Shouyi; Li, Xunde; Li, Juntao; Atwill, Edward R.
Methods in Molecular Biology. Vol. 1039 2013. p. 99-108 (Methods in Molecular Biology; Vol. 1039).Research output: Chapter in Book/Report/Conference proceeding › Chapter
}
TY - CHAP
T1 - Rapid detection of brucella spp. using loop-mediated isothermal amplification (LAMP)
AU - Chen, Shouyi
AU - Li, Xunde
AU - Li, Juntao
AU - Atwill, Edward R.
PY - 2013
Y1 - 2013
N2 - Brucella spp. are facultative intracellular bacteria that cause zoonotic disease of brucellosis worldwide. Livestock that are most vulnerable to brucellosis include cattle, goats, and pigs. Brucella spp. cause serious health problems to humans and animals and economic losses to the livestock industry. Traditional methods for detection of Brucella spp. take 48-72 h (Kumar et al., J Commun Dis 29:131-137, 1997; Barrouin-Melo et al., Res Vet Sci 83:340-346, 2007) that do not meet the food industry's need of rapid detection. Therefore, there is an urgent need of fast, specific, sensitive, and inexpensive method for diagnosing of Brucella spp. Loop-mediated isothermal amplification (LAMP) is a method to amplify nucleic acid at constant temperatures. Amplification can be detected by visual detection, fluorescent stain, turbidity, and electrophoresis. We targeted at the Brucella-specific gene omp25 and designed LAMP primers for detection of Brucella spp. Amplification of DNA with Bst DNA polymerase can be completed at 65 C in 60 min. Amplified products can be detected by SYBR Green I stain and 2.0 % agarose gel electrophoresis. The LAMP method is feasible for detection of Brucella spp. from blood and milk samples.
AB - Brucella spp. are facultative intracellular bacteria that cause zoonotic disease of brucellosis worldwide. Livestock that are most vulnerable to brucellosis include cattle, goats, and pigs. Brucella spp. cause serious health problems to humans and animals and economic losses to the livestock industry. Traditional methods for detection of Brucella spp. take 48-72 h (Kumar et al., J Commun Dis 29:131-137, 1997; Barrouin-Melo et al., Res Vet Sci 83:340-346, 2007) that do not meet the food industry's need of rapid detection. Therefore, there is an urgent need of fast, specific, sensitive, and inexpensive method for diagnosing of Brucella spp. Loop-mediated isothermal amplification (LAMP) is a method to amplify nucleic acid at constant temperatures. Amplification can be detected by visual detection, fluorescent stain, turbidity, and electrophoresis. We targeted at the Brucella-specific gene omp25 and designed LAMP primers for detection of Brucella spp. Amplification of DNA with Bst DNA polymerase can be completed at 65 C in 60 min. Amplified products can be detected by SYBR Green I stain and 2.0 % agarose gel electrophoresis. The LAMP method is feasible for detection of Brucella spp. from blood and milk samples.
KW - Blood
KW - Brucella
KW - Detection
KW - LAMP
KW - Loop-mediated isothermal amplification
KW - Milk
UR - http://www.scopus.com/inward/record.url?scp=84885571330&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=84885571330&partnerID=8YFLogxK
U2 - 10.1007/978-1-62703-535-4-8
DO - 10.1007/978-1-62703-535-4-8
M3 - Chapter
SN - 9781627035347
VL - 1039
T3 - Methods in Molecular Biology
SP - 99
EP - 108
BT - Methods in Molecular Biology
ER -