Multiplex polymerase chain reaction assays for the detection of the zearalenone chemotype of Fusarium species in white and brown rice
- Authors
- Sim, Jae Ho; Tian, Fei; Jung, Soo Yeon; Auh, Joong-Hyuck; Chun, Hyang Sook
- Issue Date
- Mar-2018
- Publisher
- ELSEVIER SCIENCE BV
- Keywords
- Fusarium species; Zearalenone chemotype; Multiplex PCR; Conventional and direct methods; Rice
- Citation
- INTERNATIONAL JOURNAL OF FOOD MICROBIOLOGY, v.269, pp 120 - 127
- Pages
- 8
- Journal Title
- INTERNATIONAL JOURNAL OF FOOD MICROBIOLOGY
- Volume
- 269
- Start Page
- 120
- End Page
- 127
- URI
- https://scholarworks.bwise.kr/cau/handle/2019.sw.cau/1060
- DOI
- 10.1016/j.ijfoodmicro.2018.02.003
- ISSN
- 0168-1605
1879-3460
- Abstract
- Early detection of the zearalenone (ZEA) chemotype of Fusarium species could be a precautionary measure for preventing ZEA contamination in rice. In this study, a multiplex polymerase chain reaction (mPCR) assay for detecting ZEA-producing fungi in rice was established using a set of four primers targeting the ZEA biosynthesis genes PKS3, PKS13, ZEB1, and ZEB2. Two mPCR approaches were used: one that amplified the DNA obtained from Fusarium isolates (conventional method) and another that directly amplified the target DNA from rice samples without time-consuming DNA isolation (direct method). The two mPCR methods showed high sensitivity in detecting ZEA-producing species, with a detection limit of 1.25 pg/mu L. of genomic DNA and 102 and 103 spores/g of white and brown rice, respectively. Both methods were specific for ZEA-producing species and gave four band patterns. The application of the two mPCR methods to 51 Fusarium isolates and 41 rice samples revealed that 31% (16 of 51) and 24% (10 of 41) of the samples were contaminated with ZEA-producing species, respectively. The mPCR results were further evaluated using high-performance liquid chromatography; in general, the two methods yielded similar results. These findings indicate that both mPCR methods are suitable for the detection of ZEA-producing Fusarium species in white and brown rice; however, the direct method yielded more rapid results.
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