Detection of HbsAg and hATIII genetically modified goats (Caprahircus) by loop-mediated isothermal amplification.
In this study, sensitive and rapid detection systems were designed using a loop-mediated isothermal amplification (LAMP) method to detect the genetically modified goats. A set of 4 primers were designed for each exogenous nucleic acids HBsAg and hATIII. The DNA samples were first amplified with the outer and inner primers and released a single-stranded DNA,of which both ends were stem-loop structure. Then one inner primer hybridized with the loop, and initiated displacement synthesis in less than 1 h. The result could be visualized by both agarose gel electrophoresis and unaided eyes directly after adding SYBR GREEN 1. The detection limit of LAMP was ten copies of target molecules, indicating that LAMP was tenfold more sensitive than the classical PCR. Furthermore, all the samples of genetically modified goats were tested positively by LAMP, and the results demonstrated that the LAMP was a rapid and sensitive method for detecting the genetically modified organism.
Key Lab of Agricultural Animal Genetics, Breeding, and Reproduction of Ministry of Education & Key Lab of Swine Genetics and Breeding of Ministry of Agriculture, Huazhong Agricultural University, Wuhan, 430070, Hubei, People's Republic of China.
Chenyu
C
Tao
Qingde
Q
Zhang
Shanli
S
Zhai
Bang
B
Liu
D000818:Animals; D030801:Animals, Genetically Modified; D000990:Antithrombin III / Q000737:chemistry / Q000235:genetics*; D001483:Base Sequence; D006041:Goats / Q000235:genetics*; D006514:Hepatitis B Surface Antigens / Q000737:chemistry / Q000235:genetics*; D006801:Humans; D008969:Molecular Sequence Data; D021141:Nucleic Acid Amplification Techniques / Q000379:methods; D012680:Sensitivity and Specificity
D006514:Hepatitis B Surface Antigens; D000990:Antithrombin III