Abstract
Toxins produced by bacteria and fungi are one of the most important factors which may cause food contamination. The study of detection methods with high sensitivity and throughput is significant for the protection of food safety. In the present study, we coupled microarray with emulsion PCR and developed a high throughput detection method. Thirteen different gene sites which encode the common toxins of several bacteria and fungi were assayed in parallel in positive and maize samples. Conventional PCR assays were carried out for comparison. The results showed that the developed microarray method had high specificity and sensitivity. Two zearalenone-related genes were investigated in one of the ten maize samples obtained with this present method. The results indicated that the emulsion based microarray detection method was developed successfully and suggested its potential application in multiple gene site detection.
Highlights
Food safety has become more and more of concern due to reported problems such as contaminated ham and cucumber
These problems could be solved by DNA microarrays, which have no requirement for different amplification products length with different genes and the sequencing information could be confirmed by hybridization, which highly increases the reliability of the method, as well as the number of detection sites is limitless for DNA microarrays
We can infer from the result that the one maize sample detected were probably contaminated by zearalenone produced by F. graminearum, because this toxin was coded mainly by the two genes PKS13 and PKS4 detected in the sample [31]
Summary
Food safety has become more and more of concern due to reported problems such as contaminated ham and cucumber. The limited number of primers has restrained its ability of parallel amplification, and only products of different length could be separated by electrophoresis These problems could be solved by DNA microarrays, which have no requirement for different amplification products length with different genes and the sequencing information could be confirmed by hybridization, which highly increases the reliability of the method, as well as the number of detection sites is limitless for DNA microarrays. PCR, BEAMing had higher efficiency with relative low concentrations of primers and good separation of the micro-droplets by introducing the magnetic beads. The side products of BEAMing were collected and labeled with fluorescence, detected by DNA microarray This BEAMing-coupled DNA microarray method was developed and validated by using to inspect fungi and bacteria which could produce toxins. The results showed that the desired gene sites were detected by the combined method and sensitively, and it revealed good application prospects
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