Scholarlink

<p>Cucumber mosaic virus (CMV), a&#xa0;member of the genus <i>Cucumovirus</i> with a&#xa0;broad host range and diverse strain variants, is one of the most prevalent plant viruses and poses a&#xa0;significant threat to global agriculture worldwide, leading to significant yield losses. The only effective way to manage plant viruses is through the use of virus-free planting material and control of vectors. Early and accurate identification of the causal agent is the most crucial step for the effective management of diseases caused by viral pathogens, for which quick, easy, and sensitive diagnostic methods are of the utmost importance. However, conventional methods, viz. the enzyme-linked immunosorbent assay (ELISA) and reverse transcription-polymerase chain reaction (RT-PCR), which are used for the detection of plant viruses, are time-consuming, costly, labor intensive and often require specialized laboratory infrastructure, limiting their utility in resource-limited settings. To address these limitations, a&#xa0;rapid and user-friendly reverse transcription–recombinase polymerase amplification–lateral flow dipstick (RT-RPA-LFD) diagnostic assay for CMV was developed. In the RT-RPA-LFD assay, the coat protein (CP) gene of CMV was successfully amplified via different templates, such as RNA, cDNA, and crude sap extracted from infected banana leaves. The assay showed a&#xa0;high sensitivity of ~0.18 pg with RNA extracted from banana leaf samples followed by cDNA (~18 fg), exhibited a&#xa0;10⁻<sup>5</sup> (1.8 pg) limit of detection with the crude sap and presented a&#xa0;clear visual signal on the LFDs. The developed assay was highly specific in the detection of CMV, and no cross-reactivity was observed with other related plant viruses infecting banana plants. The simplicity and efficacy of the RT-RPA-LFD assay offers significant advances in point-of-care disease detection, not only for crops such as banana for the selection of healthy mother plants but also for other crops that are susceptible to CMV infection.</p>

错误：搜索内容不能为空，请输入英文关键词
错误：关键词超出字数限制，请精简
高级检索

A Rapid Reverse Transcription–Recombinase Polymerase Amplification–Lateral Flow Dipstick Assay for the Detection of Cucumber Mosaic Virus in Banana Plants

  • 期刊论文

  • 发表日期：

    2025年9月30日

查看全文
  • Richa Rai,
  • Damini Diksha,
  • Pooja Bhardwaj,
  • Nishant Srivastava,
  • Nitika Gupta,
  • Susheel Kumar Sharma,
  • Virendra Kumar Baranwal

摘要

Cucumber mosaic virus (CMV), a member of the genus Cucumovirus with a broad host range and diverse strain variants, is one of the most prevalent plant viruses and poses a significant threat to global agriculture worldwide, leading to significant yield losses. The only effective way to manage plant viruses is through the use of virus-free planting material and control of vectors. Early and accurate identification of the causal agent is the most crucial step for the effective management of diseases caused by viral pathogens, for which quick, easy, and sensitive diagnostic methods are of the utmost importance. However, conventional methods, viz. the enzyme-linked immunosorbent assay (ELISA) and reverse transcription-polymerase chain reaction (RT-PCR), which are used for the detection of plant viruses, are time-consuming, costly, labor intensive and often require specialized laboratory infrastructure, limiting their utility in resource-limited settings. To address these limitations, a rapid and user-friendly reverse transcription–recombinase polymerase amplification–lateral flow dipstick (RT-RPA-LFD) diagnostic assay for CMV was developed. In the RT-RPA-LFD assay, the coat protein (CP) gene of CMV was successfully amplified via different templates, such as RNA, cDNA, and crude sap extracted from infected banana leaves. The assay showed a high sensitivity of ~0.18 pg with RNA extracted from banana leaf samples followed by cDNA (~18 fg), exhibited a 10⁻5 (1.8 pg) limit of detection with the crude sap and presented a clear visual signal on the LFDs. The developed assay was highly specific in the detection of CMV, and no cross-reactivity was observed with other related plant viruses infecting banana plants. The simplicity and efficacy of the RT-RPA-LFD assay offers significant advances in point-of-care disease detection, not only for crops such as banana for the selection of healthy mother plants but also for other crops that are susceptible to CMV infection.