Simultaneous detection of three lily viruses using Triplex IC-RT-PCR.

Area of Science:

• Plant Pathology
• Molecular Biology
• Agricultural Science

Background:

• Lilies (Lilium spp.) are susceptible to co-infection by multiple viruses, including Lily symptomless virus (LSV), Cucumber mosaic virus (CMV), and Lily mottle virus (LMoV).
• These viral co-infections lead to significant global economic losses in lily flower and bulb production, impacting both yield and quality.
• Accurate and efficient diagnostic methods are crucial for identifying these viruses and managing their spread in lily crops.

Purpose of the Study:

• To develop and validate a novel triplex immunocapture (IC) reverse transcription (RT) polymerase chain reaction (PCR) assay.
• To enable the simultaneous detection of three key lily viruses: LSV, CMV, and LMoV.
• To provide a reliable and convenient diagnostic tool for large-scale field surveys and crop health monitoring in lilies.

Main Methods:

• Development of a triplex IC-RT-PCR assay designed for the simultaneous detection of LSV, CMV, and LMoV.
• Comparison of the triplex IC-RT-PCR assay's performance against a quadruplex RT-PCR assay using field samples.
• Evaluation of diagnostic parameters including specificity, sensitivity, and agreement (κ values) for each target virus.

Main Results:

• The triplex IC-RT-PCR assay demonstrated 100% specificity for LSV, CMV, and LMoV in field samples when compared to the quadruplex RT-PCR.
• Sensitivity of the triplex IC-RT-PCR was high, reaching 99.4% for LSV, 81.4% for CMV, and 98.7% for LMoV.
• Excellent agreement (κ values of 0.968, 0.844, and 0.984 for LSV, CMV, and LMoV, respectively) was observed between the triplex IC-RT-PCR and the quadruplex RT-PCR methods.

Conclusions:

• This study reports the first successful development of a triplex IC-RT-PCR assay for the simultaneous detection of LSV, CMV, and LMoV.
• The developed assay is highly specific and sensitive, offering a reliable alternative to existing detection methods.
• The triplex IC-RT-PCR assay is a convenient and effective tool suitable for routine use in large-scale lily field surveys and crop health management programs.