[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.
National Institute of Health Sciences, 1-18-1, Kamiyoga, Setagaya-ku, Tokyo 158-8501, Japan.
Hiroshi
H
Akiyama
Kazuto
K
Isuzugawa
Naoki
N
Harikai
Hiroko
H
Watanabe
Ken
K
Iijima
Hirohito
H
Yamakawa
Yamato
Y
Mizuguchi
Reiji
R
Yoshikawa
Miho
M
Yamamoto
Hidetaka
H
Sato
Masatoshi
M
Watai
Fumihiro
F
Arakawa
Takeshi
T
Ogasawara
Rikuka
R
Nishihara
Hisashi
H
Kato
Atsushi
A
Yamauchi
Yoshihisa
Y
Takahata
Fumiki
F
Morimatsu
Shinichi
S
Mamegoshi
Shiroo
S
Muraoka
Tsutomu
T
Honjoh
Takahiro
T
Watanabe
Kozue
K
Sakata
Tomoaki
T
Imamura
Masatake
M
Toyoda
Rieko
R
Matsuda
Tamio
T
Maitani
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