Schmidt-Heydt M, Richter W, Michulec M, Buttinger G, Geisen R
Max Rubner Institute-Federal Research Centre of Nutrition and Food, Karlsruhe, Germany.
Food Addit Contam Part A Chem Anal Control Expo Risk Assess. 2008 Aug;25(8):989-96. doi: 10.1080/02652030801961305.
Based on the sequence of the ochratoxin A polyketide synthase gene (otapksPV), a polymerase chain reaction (PCR) system for the specific detection of Penicillium verrucosum in wheat has been developed. In a further approach, a real-time PCR system has been applied to determine the growth kinetics of P. verrucosum in wheat at cell numbers above 10(3) colony-forming units (cfu) ml(-1). The data obtained by real-time PCR correlated well with the data obtained by the plate count technique. For this purpose, the DNA was isolated directly from contaminated wheat without any further enrichment step. In a reverse transcriptase real-time PCR, the expression of the otapksPV gene in wheat was detected 22 days after inoculation and storage at ambient temperature. Reasonable amounts of ochratoxin A, however, could not be detected before day 30. This early activation of ochratoxin A related genes was confirmed by microarray analysis.
基于赭曲霉毒素A聚酮合酶基因（otapksPV）的序列，开发了一种用于特异性检测小麦中疣孢青霉的聚合酶链反应（PCR）系统。在进一步的研究中，应用实时PCR系统来确定小麦中疣孢青霉在细胞数高于10³菌落形成单位（cfu）ml⁻¹时的生长动力学。实时PCR获得的数据与平板计数技术获得的数据相关性良好。为此，直接从受污染的小麦中分离DNA，无需任何进一步的富集步骤。在逆转录实时PCR中，接种并在室温下储存22天后检测到小麦中otapksPV基因的表达。然而，在第30天之前未检测到适量的赭曲霉毒素A。通过微阵列分析证实了赭曲霉毒素A相关基因的这种早期激活。
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