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<p>Potato virus Y (PVY) is a major threat to global potato production, causing significant yield losses. Being seed borne and aphid transmitted virus it has significant potential to spread to new geographical locations. To limit its spread and protect crops from heavy losses, a rapid, highly sensitive, and specific detection method is essential. Current detection of viruses is mainly based on enzyme-linked immunosorbent assay (ELISA) and polymerase chain reaction (PCR). However these methods are labour-intensive, demand expensive equipment, technical skill, and are often time-consuming. Recently, isothermal amplification, especially loop-mediated isothermal amplification (LAMP), has been demonstrated as a simple, rapid, highly sensitive and specific detection technique making it a promising alternative for the detection of various pathogens. In the present study, we have optimized RT-LAMP assay for detection of PVY with high sensitivity and specificity at 60&#xa0;°C in just 30&#xa0;min. Six primer sets were designed targeting highly conserved region of PVY. Specificity of the assay was determined using six non target <i>Potyviruses</i> (BCMV, BCMNV, ClYVV, SMV, PVA and PVM) and no cross reactivity was detected. Furthermore, RT-LAMP assay sucessfully detected PVY upto 10<sup>–14</sup> dilution starting from 100&#xa0;ng/μL and was found 10,000 more sensitive compared to RT-PCR. For practical field applications, the RT-LAMP assay was designed to detect PVY directly from crude sap, eliminating the need for complex laboratory equipment. A colorimetric assay was also incorporated for easy visualization of results, enabling rapid and accurate detection of PVY in both laboratory and field settings. This developed RT-LAMP assay is simple, fast, highly sensitive, and specific, making it an ideal tool for PVY detection in diverse environments.</p>

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A Rapid and Field-Deployable RT-LAMP Assay for Reliable Detection of Potato Virus Y (PVY)

  • 期刊论文

  • 发表日期：

    2025年4月29日

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  • Shahjahan Rashid,
  • Sumiah Wani,
  • Nulevino Iralu,
  • Gowhar Ali,
  • Aflaq Hamid

摘要

Potato virus Y (PVY) is a major threat to global potato production, causing significant yield losses. Being seed borne and aphid transmitted virus it has significant potential to spread to new geographical locations. To limit its spread and protect crops from heavy losses, a rapid, highly sensitive, and specific detection method is essential. Current detection of viruses is mainly based on enzyme-linked immunosorbent assay (ELISA) and polymerase chain reaction (PCR). However these methods are labour-intensive, demand expensive equipment, technical skill, and are often time-consuming. Recently, isothermal amplification, especially loop-mediated isothermal amplification (LAMP), has been demonstrated as a simple, rapid, highly sensitive and specific detection technique making it a promising alternative for the detection of various pathogens. In the present study, we have optimized RT-LAMP assay for detection of PVY with high sensitivity and specificity at 60 °C in just 30 min. Six primer sets were designed targeting highly conserved region of PVY. Specificity of the assay was determined using six non target Potyviruses (BCMV, BCMNV, ClYVV, SMV, PVA and PVM) and no cross reactivity was detected. Furthermore, RT-LAMP assay sucessfully detected PVY upto 10–14 dilution starting from 100 ng/μL and was found 10,000 more sensitive compared to RT-PCR. For practical field applications, the RT-LAMP assay was designed to detect PVY directly from crude sap, eliminating the need for complex laboratory equipment. A colorimetric assay was also incorporated for easy visualization of results, enabling rapid and accurate detection of PVY in both laboratory and field settings. This developed RT-LAMP assay is simple, fast, highly sensitive, and specific, making it an ideal tool for PVY detection in diverse environments.