Loop-mediated isothermal amplification technique: principle, development and wide application in food safety

2020 ◽  
Vol 12 (46) ◽  
pp. 5551-5561
Author(s):  
Tianzeng Huang ◽  
Linzhi Li ◽  
Xing Liu ◽  
Qi Chen ◽  
Xueen Fang ◽  
...  
Keyword(s):  
Food Safety ◽  
Gene Amplification ◽  
Foodborne Pathogens ◽  
High Specificity ◽  
Isothermal Amplification ◽  
Isothermal Conditions ◽  
Loop Mediated Isothermal Amplification ◽  
Novel Gene ◽  
Amplification Method ◽  
Amplification Technique

LAMP is a relatively novel gene amplification method under isothermal conditions with rapidity, and high specificity. It is widely applied in the field of food safety, such as in the detection of foodborne pathogens, GM, OP pesticides and so on

Loop-Mediated Isothermal Amplification Method for Rapid Detection of Shigella dysenteriae

2011 ◽  
Vol 140 ◽  
pp. 369-373 ◽  
Author(s):  
Qing Ping Zhong ◽  
Li Wang ◽  
Bin Wang ◽  
Hong Yuan Chen
Keyword(s):  
Isothermal Condition ◽  
Lamp Assay ◽  
High Specificity ◽  
Isothermal Amplification ◽  
Shigella Dysenteriae ◽  
Lamp Method ◽  
Loop Mediated Isothermal Amplification ◽  
Amplification Method ◽  
Target Dna ◽  
Pcr Method

The study was aimed to develop a loop-mediated isothermal amplification (LAMP) method which amplifies DNA with high specificity and rapidity for the detection of Shigella dysenteriae. A set of four primers was designed for recognizing six distinct sequences on the target ipaH of S. dysenteriae. By the method, the target DNA was amplified within 1h under isothermal condition at 65 °C. The sensitivities of the LAMP for detecting pure culture and genomic DNA were 1.04 CFU/ml and 1.06 fg/μl, while the sensitivities of PCR method were 1.04×102 CFU/ml and 1.06 pg/μl. Furthermore, the LAMP assay was examined for its ability to detect S. dysenteriae in artificially contaminated lettuce sample, the detection limits of this LAMP assay and the PCR method were 4.60 CFU/g and 4.60×102 CFU/g, respectively.

scholarly journals Efficient and Specific Detection ofSalmonellain Food Samples Using astn-Based Loop-Mediated Isothermal Amplification Method

2015 ◽  
Vol 2015 ◽  
pp. 1-7 ◽  
Author(s):  
Mevaree Srisawat ◽  
Watanalai Panbangred
Keyword(s):  
Lamp Assay ◽  
High Sensitivity ◽  
High Specificity ◽  
Culture Method ◽  
Food Samples ◽  
Isothermal Amplification ◽  
Lamp Method ◽  
Loop Mediated Isothermal Amplification ◽  
Amplification Method ◽  
Pure Cultures

TheSalmonellaenterotoxin (stn) gene exhibits high homology amongS. entericaserovars andS. bongori. A set of 6 specific primers targeting thestngene were designed for detection ofSalmonellaspp. using the loop-mediated isothermal amplification (LAMP) method. The primers amplified target sequences in all 102 strains of 87 serovars ofSalmonellatested and no products were detected in 57 non-Salmonellastrains. The detection limit in pure cultures was 5 fg DNA/reaction when amplified at 65°C for 25 min. The LAMP assay could detectSalmonellain artificially contaminated food samples as low as 220 cells/g of food without a preenrichment step. However, the sensitivity was increased 100-fold (~2 cells/g) following 5 hr preenrichment at 35°C. The LAMP technique, with a preenrichment step for 5 and 16 hr, was shown to give 100% specificity with food samples compared to the reference culture method in which 67 out of 90 food samples gave positive results. Different food matrixes did not interfere with LAMP detection which employed a simple boiling method for DNA template preparation. The results indicate that the LAMP method, targeting thestngene, has great potential for detection ofSalmonellain food samples with both high specificity and high sensitivity.

scholarly journals P02.06 A Novel Loop-Mediated Isothermal Amplification Method for Robust Detection of EGFR Mutations

2021 ◽  
Vol 16 (3) ◽  
pp. S248-S249
Author(s):  
Y. Saito ◽  
A. Matsui ◽  
S. Michiyuki ◽  
Y. Yamauchi ◽  
N. Takahashi ◽  
...  
Keyword(s):  
Isothermal Amplification ◽  
Egfr Mutations ◽  
Robust Detection ◽  
Loop Mediated Isothermal Amplification ◽  
Amplification Method

scholarly journals Sensitive and Rapid Detection of Edwardsiellosis in Fish by a Loop-Mediated Isothermal Amplification Method

2004 ◽  
Vol 70 (1) ◽  
pp. 621-624 ◽  
Author(s):  
Ram Savan ◽  
Arisa Igarashi ◽  
Satoru Matsuoka ◽  
Masahiro Sakai
Keyword(s):  
Rapid Detection ◽  
Japanese Flounder ◽  
Isothermal Amplification ◽  
Edwardsiella Tarda ◽  
Fish Disease ◽  
Fish Pathogen ◽  
Loop Mediated Isothermal Amplification ◽  
Diagnostic Protocol ◽  
Amplification Method ◽  
Hemolysin Gene

ABSTRACT Here we report a rapid and sensitive method (using loop-mediated isothermal amplification [LAMP]) for the diagnosis of edwardsiellosis, a fish disease caused by Edwardsiella tarda, in Japanese flounder. A set of four primers was designed, and conditions for the detection were optimized for the detection of E. tarda in 45 min at 65°C. No amplification of the target hemolysin gene was detected in other related bacteria. When the LAMP primers were used, detection of edwardsiellosis in infected Japanese flounder kidney, and spleen and seawater cultures was possible. We have developed a rapid and sensitive diagnostic protocol for edwardsiellosis detection in fish. This is the first report of the application of LAMP for the diagnosis of a fish pathogen.

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