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Bangladesh Agricultural University (BAU)

Publish Date: 28 Apr 2026

Validation of Loop-Mediated Isothermal Amplification (LAMP) for Rapid Diagnosis of Fusarium oxysporum f. sp. cubense Tropical Race 4 (Foc TR4) causing Fusarium Wilt of Banana

Dr. Md. Rashidul Islam

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Banana is one of the important tropical fruits which is grown as cash crop by the smallholder farmers and is an important source of nutrients for the people in Bangladesh. Fusarium wilt (FW) of banana caused by Fusarium oxysporum f. sp. cubense tropical race 4 (Foc TR4) is one of the devastating diseases incurred huge losses to the farm families globally. In this study, the existence and prevalence of Foc TR4 were confirmed by PCR and Loop-Mediated Isothermal Amplification (LAMP) based rapid diagnosis was validated. A total of 354 Foc isolates were obtained from a total of 351 FW infected banana plant samples (pseudostem/rhizome) of different varieties collected from 34 districts across the country. Diagnosis of 354 single spore purified Foc isolates by PCR showed that a total of 212 Foc isolates belongs to Foc TR4 (VCG group 01213/16) that are obtained from different varieties of 19 districts. LAMP was validated for the rapid diagnosis (within an hour) of Foc TR4 in both laboratory and field conditions. The specificity of LAMP assay was evaluated using banana samples previously identified as Foc TR4 by PCR, along with positive and negative controls. All TR4-inoculated samples, including pseudostem (A1) and rhizome (A2), exhibited rapid and distinct fluorescence amplification curves, confirming the presence of Foc TR4 DNA. Positive DNA controls (A3 and A4) also showed strong amplification, whereas the negative control containing nuclease-free water (A6) produced no amplification, demonstrating high assay specificity. The sensitivity of the assay was further assessed using diluted DNA samples (A3-A5), which generated amplification curves corresponding to their DNA concentrations, indicating the assay’s ability to detect low-copy DNA reliably in the presence of background plant DNA. These results collectively revealed the use of LAMP for phytosanitary and on-site rapid diagnosis of Foc TR4 to prevent its spread in the country.

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