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A single polyprobe for detecting simultaneously eight pospiviroids infecting ornamentals and vegetables
Enza Maria Torchetti1, Beatriz Navarro, Francesco Di Serio
1Istituto di Virologia Vegetale del CNR, UOS Bari, Via Amendola 165/A, 70126 Bari, Italy.
Journal of Virological Methods
|September 1, 2012
Summary
A new digoxigenin-labeled polyprobe (POSPIprobe) effectively detects multiple Pospiviroid species in plants. This tool aids in rapid identification for quarantine and certification programs.
Area of Science:
- Plant pathology
- Molecular biology
- Virology
Background:
- Viroids of the genus Pospiviroid are spreading in European ornamental and vegetable crops.
- Current detection methods require improvement in speed, efficiency, and sensitivity.
Purpose of the Study:
- To develop a sensitive and efficient detection method for Pospiviroid species.
- To create a broad-spectrum polyprobe for Pospiviroid detection.
Main Methods:
- Bioinformatic analysis of Pospiviroid sequences.
- Development of a digoxigenin-labeled polyprobe (POSPIprobe).
- Testing the POSPIprobe using dot-blot and Northern-blot hybridization assays.
Main Results:
- The POSPIprobe successfully detected eight Pospiviroid species.
- Detection limits were comparable to single probes, ranging from 5(-2) to 5(-4) and 5(-1) to 5(-3) depending on nucleic acid extraction method.
- Validation with 68 field samples confirmed the polyprobe's efficacy.
Conclusions:
- The POSPIprobe is a valuable tool for the rapid and sensitive detection of Pospiviroid infections.
- It holds significant potential for use in plant quarantine, certification, and survey programs.
- The probe is expected to detect additional Pospiviroid species not included in the initial study.
