Improved multiplex reverse transcription-polymerase chain reaction to detect and identify five tospovirus species simultaneously

Abstract

We improved a one-step multiplex reverse transcription-polymerase chain reaction (multi-PCR) method previously published for the simultaneous identification of five tospovirus species that mainly occur on Solanaceae and Compositae plants. The multi-PCR system is composed of a universal degenerate primer and five virus-species-specific primers that amplify bands unique to Iris yellow spot virus (837 bp), Tomato spotted wilt virus (709 bp), Impatiens necrotic spot virus (589 bp), Chrysanthemum stem necrosis virus (485 bp), and Capsicum chlorosis virus (366 bp). This technique is potentially useful for epidemiological studies of several tospoviruses that have recently been detected in previously uninfected fields of ornamental crops.