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目的了解成都地区市售辣椒粉受霉菌污染的状况,探讨裸磁珠-多重实时荧光定量PCR方法快速定量检测霉菌数量的实用性。方法于成都市区随机选取农贸市场,采集辣椒样品40份,按照国标GB4789.15-2010对样品进行霉菌计数,纯化后以镜下特征及菌落特征进行菌种鉴定及分类计数,同时采用裸磁珠-实时荧光PCR方法检测霉菌数量,通过与传统方法比较,探讨裸磁珠-实时荧光PCR方法的实用性。结果所采集样品的霉菌污染均数达到了5.1×103CFU/g;对曲霉属、青霉属及镰刀菌属分类计数,裸磁珠-多重实时荧光定量PCR的检测法计数结果几何均数分别为6.1×108、4.0×101及2.9×106CFU/g,此结果与传统培养计数法2.0×103、1.1×102及2.1×101CFU/g相比有显著增加,两者结果具有统计学差异。结论成都市售辣椒粉受霉菌污染严重,对消费者健康构成较大威胁。相比传统培养法,裸磁珠-多重实时荧光定量PCR方法节约时间、灵敏度高且受稀释等操作的影响较小、受样本成分及样本中其他霉菌干扰较小,具有很高的应用价值。
Objective To investigate the status of fungal contamination of chili powder on the market in Chengdu, and to explore the practicability of using naked magnetic beads - multiplex real-time quantitative PCR to detect the quantity of mold rapidly and quantitatively. Methods 40 farmer’s markets were randomly selected in the urban area of Chengdu. 40 samples of peppers were collected. The molds were counted according to the national standard GB4789.15-2010. After purification, the bacteria were identified by microscopic features and colony characteristics, The method of bead-real-time fluorescence PCR was used to detect the number of molds, and the practicability of naked magnetic bead-real-time fluorescence PCR was discussed by comparison with traditional methods. Results The average number of mold contamination reached 5.1 × 103CFU / g. The counts of Aspergillus, Penicillium and Fusarium were classified. The geometric mean of count of naked magnetic beads - multiplex real-time PCR was 6.1 × 108, 4.0 × 101 and 2.9 × 106CFU / g, respectively. Compared with the traditional culture counting method, the results showed a significant increase compared with the traditional culture counting method of 2.0 × 103, 1.1 × 102 and 2.1 × 101CFU / g. Conclusion The chili powder sold in Chengdu is seriously polluted by mold and poses a great threat to the health of consumers. Compared with the traditional culture method, the naked magnetic bead-multiplex real-time PCR method has the advantages of saving time, high sensitivity and being less affected by dilution and other operations, and is less disturbed by the sample components and other molds in the sample, which has high application value.