Detection of HbsAg and hATIII genetically modified goats (Caprahircus) by loop-mediated isothermal amplification

Mol Biol Rep. 2013 Nov;40(11):6177-82. doi: 10.1007/s11033-013-2728-7. Epub 2013 Sep 22.

Abstract

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Animals, Genetically Modified
  • Antithrombin III / chemistry
  • Antithrombin III / genetics*
  • Base Sequence
  • Goats / genetics*
  • Hepatitis B Surface Antigens / chemistry
  • Hepatitis B Surface Antigens / genetics*
  • Humans
  • Molecular Sequence Data
  • Nucleic Acid Amplification Techniques* / methods
  • Sensitivity and Specificity

Substances

  • Hepatitis B Surface Antigens
  • Antithrombin III