Abstract
Rice false smut is a worldwide devastating rice disease infected by the fungal pathogen Villosiclava virens. Ustiloxin A (UA) and ustiloxin B (UB), cyclopeptide mycotoxins, were the major ustiloxins isolated from the rice false smut balls (FSBs) that formed in the pathogen-infected rice spikelets. Based on the specific monoclonal antibodies (mAbs) 2D3G5 and 1B5A10, respectively, against UA and UB, the lateral flow immunoassays (LFIAs) were developed, and the indicator ranges for UA and UB both were 50–100 ng/mL. The cross-reactivities of UB for UA LFIA, and UA for UB LFIA were 5% and 20%, respectively, which were consistent with the icELISA results reported previously. Even at 50,000 ng/mL, none of other commonly existent metabolites in rice samples caused noticeable inhibition. The LFIAs were used for determination of UA and UB contents in rice FSBs and rice grains, and the results were agreeable with those by HPLC and icELISA. There was no change in the sensitivity of either dipstick stored at 4 °C after at least three months. The developed LFIA has specificity and sensitivity for detecting UA and UB as well as simplicity to use. It will be a potential point-of-care device for rapid evaluation of the rice samples contaminated by UA and UB.
Highlights
Rice false smut is a worldwide destructive rice disease caused by the ascomycete fungus Villosiclava virens (Nakata) Tanaka and Tanaka [1] in most rice-growing areas such as China, India, Japan and USA over the past few years [2,3,4,5]
For lateral flow immunoassays (LFIAs) of ustiloxin B (UB), there was no change in the sensitivity (50–100 ng/mL of UB) after a storage of six months at 4 ◦ C
The LFIAs for Ustiloxin A (UA) and UB were developed based on the monoclonal antibodies (mAbs) 2D3G5 and 1B5A10 respectively
Summary
Rice false smut is a worldwide destructive rice disease caused by the ascomycete fungus Villosiclava virens (Nakata) Tanaka and Tanaka (anamorph: Ustilaginoidea virens Takahashi) [1] in most rice-growing areas such as China, India, Japan and USA over the past few years [2,3,4,5]. Compared with the conventional ELISA, LFIA is an economic method with a simple preparation, and the results can be read by naked eyes within 3–10 min, which is highly suitable as a sample preparation, and the results can be read by naked eyes within 3–10 min, which is highly point‐of‐care diagnostic device without using expensive instrumentation [30]. A and easy‐to‐use diagnostic device for rapid evaluation of the ustiloxin contamination in rice samples convenient and easy-to-use diagnostic device for rapid evaluation of the ustiloxin contamination in rice at the point‐of‐care is still lacking To develop such a LFIA, the antibody has been considered as the samples at the point-of-care is still lacking. Qualitative and semi-quantitative analysis of UA and UB contents in rice samples were evaluated
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