Study of PCR Detection Methods for Genetically Modified Soybeans with Reference Molecules
HSU-YANG LIN1, HSIU-AN WEI1, Fu-Pang Lin2 AND DANIEL YANG-CHIH SHIH1*
1. Bureau of Food and Drug Analysis, Department of Health, Executive Yuan, 161-2, Kunyang St., Taipei 115, Taiwan, R.O.C.
2. Institute of Bisocience and Biotechnology, National Taiwan Ocean University, 2, Pei-Ning Rd., Keelung 202, Taiwan, R.O.C.
(Received: August 1, 2005; Accepted: February 21, 2006)
ABSTRACT
The study of detection methods for genetically modified (GM) soy using certified reference material (CRM) and novel references molecules was operated on LightCycler real-time PCR machines system. The test results of this study demonstrated the methods used to be applicable to the specific quantitation of one line of GM soy. Independent repeat tests for 5, 2 and 1% CRM were 17, 11 and 11, and the test results mean ± SD were 4.89 ± 0.45, 1.98 ± 0.63 and 1.09 ± 0.05, respectively. Series 6 repeat test by use of CRM and references molecules, the results of CRM test were 5.45 ± 0.32, 2.42 ± 0.13 and 1.20 ± 0.09 (mean ± SD), and 4.83 ± 0.45, 2.09 ± 0.12 and 1.08 ± 0.10 for references molecules testing, to 5, 2, and 1% GM content, respectively. Further, soymilk was not detected by the ELISA method at OD450 when boiled under 70°C, 1~3 min but high to 100°C was detected by PCR method. Over 10 min under 121°C, DNA highly degradation detection was more difficult. Testing results should help support the practical detection for the GM soy.
Key word: real-time PCR, PCR, ELISA, reference molecules, GMO, soybeans, quantification