Author
WATANABE, TAKAHIRO - JAPAN INSTITUTE | |
AKIYAMA, HIROSHI - JAPAN INSTITUTE | |
Maleki, Soheila | |
YAMAKAW, HIROHITO - JAPAN INSTITUTE | |
IIJIMA, KEN - JAPAN INSTITUTE | |
YAMAZAKI, FUMINORI - JAPAN INSTITUTE | |
MATSUMOTO, TAKASHI - JAPAN INSTITUTE | |
FUTO, SATOSHI - JAPAN INSTITUTE | |
ARAKAWA, FUMIHIRO - JAPAN INSTITUTE | |
WATAI, MASATOSHI - JAPAN INSTITUTE |
Submitted to: Journal of Food Biochemistry
Publication Type: Peer Reviewed Journal Publication Acceptance Date: 8/2/2005 Publication Date: 1/3/2006 Citation: Watanabe, T., Akiyama, H., Maleki, S.J., Yamakaw, H., Iijima, K., Yamazaki, F., Matsumoto, T., Futo, S., Arakawa, F., Watai, M. 2006. A specific qualitative detection method for peanut (arachis hypogagea) in foods using polymerase chain reaction. Journal of Food Biochemistry. 215-233. Interpretive Summary: A qualitative method for detection of peanuts in foods using polymerase chain reaction (PCR) was developed. A universal DNA fragement pair (CP 03-5 /CP 03-3) was designed to confirm the validity of the DNAs for PCR. In addition, for the specific detection of peanuts with high sensitivity, the primer pair agg 04-5 /agg 05-3 was designed to detect the gene encoding the peanut agglutinin precursor. The primer pair specifically generates a 95-bp amplified fragment from peanut genomic DNA. Five hundred femto grams of peanut genomic DNA can be detected using the established method. The same qualitative results were obtained from both model processed and non-processed food samples containing 0.001, 0.01 and 0.1% peanut. Moreover, it was shown that the trace amount of peanut in the commercial food products could be qualitatively detected using this method. The reproducibility and applicability of the proposed methods were verified in a six-laboratory collaborative study. Technical Abstract: A qualitative method for detection of peanuts in foods using polymerase chain reaction was developed. A universal primer pair CP 03-5 /CP 03-3 was designed to confirm the validity of the DNAs for PCR. The plant-specific amplified fragments were detected from 13 kinds of plants using the universal primer pair. In addition, for the specific detection of peanuts with high sensitivity, the primer pair agg 04-5 /agg 05-3 was designed to detect the gene encoding the peanut agglutinin precursor. The primer pair specifically generates a 95-bp amplified fragment from peanut genomic DNA. Five hundred femto grams of peanut genomic DNA can be detected using the established method. The same qualitative results were obtained from both model processed and nonprocessed food samples containing 0.001, 0.01 and 0.1% peanut. Moreover, it was shown that the trace amount of peanut in the commercial food products could be qualitatively detected using this method. The reproducibility and applicability of the proposed methods were verified in a six-laboratory collaborative study. |