14982nas a2200145 45000080041000002450131000412100069001722600008002413000011002494900006002605201438100266100003314647700003014680856012614710 2025 eng d00aDevelopment of multiplex PCR assay for detection of mycotoxigenic Fusarium species contamination in stored maize grains, India0 aDevelopment of multiplex PCR assay for detection of mycotoxigeni cMAR a1002400 v63 a

This investigation aimed to improve the detection of key mycotoxigenic Fusarium species to address agricultural biosecurity and food safety threats in stored maize grains in India. We developed a multiplex polymerase chain reaction (mPCR) assay for the selective detection of Fusarium species producing mycotoxinfumonisin B1 (FB1), zearalenone (ZEA), and deoxynivalenol (DON) using their biosynthetic pathway genes viz., FUM1, FUM13, PKS4, PKS13 and TRI13, TRI7, respectively. The mPCR assay demonstrated a limit of detection (LOD) ranging from 1 × 103 and 1 × 102 spores/g maize grain in both naturally contaminated and Fusarium spores spiked samples, respectively. Maize grain samples were also evaluated for the assessment of mycotoxigenic Fusarium species contamination by mPCR which resulted in A total of 53 maize grain samples (75 %) positive for mycotoxin chemotypes upon mPCR analysis. Among these, 40 sample tested positive for FB1, 39 for ZEA and 25 for DON. The levels of contamination ranged from 4 to 6456 µg/kg for FB1, 30–2192 µg/kg for ZEA and 5–1713 µg/kg for DON as determined by high-performance liquid chromatography (HPLC). Furthermore, the developed mPCR results exhibited a significant correlation (>95 %) with HPLC data confirming this assays reliability. This finding infers developed mPCR assay is an expeditious, cost-effective, sensitive and selective tool for detecting toxigenic Fusarium species. This method can play a crucial role in assessing food safety and public health, particularly in regions where maize contamination by mycotoxins is a significant concern.

1 aNavale, Vishwambar, Digambar1 aVamkudoth, Koteswara, Rao uhttps://library.ncl.res.in/content/development-multiplex-pcr-assay-detection-mycotoxigenic-fusarium-species-contamination