[Inter-laboratory evaluation studies for development of notified ELISA methods for allergic substances (wheat)].
Extracts of sausage, sauce, pasta sauce, fish paste and cereal spiked with wheat standard protein at a level of 5-20 ng/mL as sample solutions were analyzed in replicate in 10 laboratories. Coefficients of variation (CVs) of both ELISA methods using a Wheat Protein ELISA Kit (Gliadin kit) and a FASTKIT Wheat ELISA Kit (Wheat ELISA kit) were mostly below 10%. Mean recoveries of the wheat standard protein from the food extracts were over 40% in the two ELISA methods except those from cereal extract determined using the Wheat ELISA kit. Repeatability relative standard deviations of wheat standard protein in the five food extracts were in the ranges of 16-26.9% and 3.7-36.2% for the Gliadin kit and the Wheat ELISA kit, respectively. Reproducibility relative standard deviations of wheat standard protein in the five food extracts were 21.6-38.5%, 29.7-53.8% for the Gliadin kit and the Wheat ELISA kit, respectively. The recoveries of wheat standard protein from the cereal extract were improved by the increasing the amount of antibody coated on the plate in the Wheat ELISA kit. The detection limits of both ELISA methods were 1 ng/mL in sample solutions. These results suggested that the notified ELISA methods are reliable and reproducible for the inspection of wheat protein levels in extracts of sausage, sauce, pasta sauce, fish paste and cereal.
Akiyama
Hiroshi
H
National Institute of Health Sciences, 1-18-1, Kamiyoga, Setagaya-ku, Tokyo 158-8501, Japan.
Isuzugawa
Kazuto
K
Harikai
Naoki
N
Watanabe
Hiroko
H
Iijima
Ken
K
Yamakawa
Hirohito
H
Mizuguchi
Yamato
Y
Yoshikawa
Reiji
R
Yamamoto
Miho
M
Sato
Hidetaka
H
Watai
Masatoshi
M
Arakawa
Fumihiro
F
Ogasawara
Takeshi
T
Nishihara
Rikuka
R
Kato
Hisashi
H
Yamauchi
Atsushi
A
Takahata
Yoshihisa
Y
Morimatsu
Fumiki
F
Mamegoshi
Shinichi
S
Muraoka
Shiroo
S
Honjoh
Tsutomu
T
Watanabe
Takahiro
T
Sakata
Kozue
K
Imamura
Tomoaki
T
Toyoda
Masatake
M
Matsuda
Rieko
R
Maitani
Tamio
T
D000485:Allergens / Q000032:analysis*; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D005504:Food Analysis / Q000379:methods*; D015337:Multicenter Studies as Topic; D010940:Plant Proteins / Q000032:analysis*; D011933:Reagent Kits, Diagnostic / Q000592:standards*; D015203:Reproducibility of Results; D014908:Triticum
D000485:Allergens; D010940:Plant Proteins; D011933:Reagent Kits, Diagnostic