CBH351玉米在筛选PCR分析中假阳性反应的调查。

Investigation of false-positive reactions for CBH351 maize in screening PCR analysis.

作者信息

Monma Kimio, Moriuchi Rie, Sagi Naoki, Ichikawa Hisatsugu, Satoh Kazue, Tobe Takashi, Kamata Kunihiro

机构信息

Tokyo Metropolitan Institute of Public Health, 3-24-1, Hyakunin-cho, Shinjuku-ku, Tokyo 169-0073, Japan.

出版信息

Shokuhin Eiseigaku Zasshi. 2006 Feb;47(1):9-14. doi: 10.3358/shokueishi.47.9.

DOI:10.3358/shokueishi.47.9

PMID:16619851

Abstract

Examination for CBH351 maize was conducted by the qualitative polymerase chain reaction (PCR) method in maize grain and maize processed foods obtained in the Tokyo area. The numbers of samples possibly positive in the screening test were 7 of 22 (31.8%) for maize grain samples, 4 of 14 (28.6%) for semi-processed foods, 11 of 30 (36.7%) for canned products, 3 of 30 (10.0%) for maize snacks, 3 of 4 (75%) for tacos and 1 of 3 (33.3%) for tortillas. However, CBH351 maize was not detected in the confirmation test. Therefore, the results of the screening test were false-positive. Since the reaction might have been caused by the base sequences of the 3'-end of primers CaM03-5' and CBH02-3' used in the screening test, a new primer pair was designed. The PCR products obtained with the new primer pair TMC2-5'--TMS2-3' were specific for CBH351 and were not obtained with barley, wheat, rice, RRS, Bt11, or Event176. Thus, the new primer pair shows high specificity. CBH351 maize was detected from samples containing at least 0.05% CBH 351 maize DNA by using this primer pair.

摘要

采用定性聚合酶链反应（PCR）方法，对东京地区获取的玉米籽粒及玉米加工食品进行了CBH351玉米检测。在筛选试验中，玉米籽粒样本22份中有7份（31.8%）可能呈阳性，半成品食品14份中有4份（28.6%），罐装产品30份中有11份（36.7%），玉米零食30份中有3份（10.0%），墨西哥玉米卷4份中有3份（75%），玉米粉圆饼3份中有1份（33.3%）。然而，在确证试验中未检测到CBH351玉米。因此，筛选试验结果为假阳性。由于该反应可能是由筛选试验中使用的引物CaM03 - 5'和CBH02 - 3'的3'端碱基序列引起的，因此设计了一对新引物。用新引物对TMC2 - 5'--TMS2 - 3'获得的PCR产物对CBH351具有特异性，而大麦、小麦、水稻、RRS、Bt11或Event176则未获得该产物。因此，新引物对具有高度特异性。使用该引物对可从含有至少0.05% CBH 351玉米DNA的样本中检测到CBH351玉米。