@article {50009, title = {Development of multiplex PCR assay for detection of mycotoxigenic Fusarium species contamination in stored maize grains, India}, journal = {The Microbe}, volume = {6}, year = {2025}, month = {MAR}, pages = {100240}, type = {Article}, abstract = {

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\ mycotoxin,\ fumonisin\ 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 {\texttimes} 103\ and 1 {\texttimes} 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\ mycotoxinchemotypes\ 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 {\textmu}g/kg for FB1, 30{\textendash}2192 {\textmu}g/kg for ZEA and 5{\textendash}1713 {\textmu}g/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.

}, doi = {https://doi.org/10.1016/j.microb.2025.100240}, author = {Navale, Vishwambar Digambar and Vamkudoth, Koteswara Rao} }