Using RT-PCR for plant RNA virus detection.

Abstract

Abstract Plants can be infected by a wide range of viruses that often cause important agronomic, economic and social impacts. Detection of viruses at premature stages of infection is crucial to allow control viral diseases and reduce economic losses. For that reason, the use of rapid, sensitive and accurate detection methods is recommended. Molecular techniques have revolutionized the way of plant virus detection and identification. Retrotranscription (RT)-polymerase chain reaction (PCR) is the 'gold standard' method for virus detection in most laboratories worldwide because of its sensitivity, specificity, simplicity and rapidity. Since its discovery, many versions and continuous improvements of PCR have been developed. Nested and co-operational RT-PCR achieve high sensitivity, decreasing contamination risks and multiplex RT-PCR offer the possibility to detect several targets in one assay. The development of real-time RT-PCR completely revolutionized the way of virus detection and quantitation, contributing enormously to the control of plant virus diseases. Some novel molecular methods such as nucleic acids arrays, isothermal amplification, massive sequencing and biosensors could be applied in the future in most diagnostic laboratories. However, the use of validated and integrated protocols that include molecular techniques combined with serological or biological methods will increase the accuracy and reliability of virus detection. In this review, the main RT-PCR-based methods used for plant virus detection are described and the advantages and drawbacks of each of them are discussed.