Detection of Tospovirus species by RT-PCR of the N gene and restriction enzymes digestion of the products.

R. A. DEWEY; L SEMORILE; O GRAU

JOURNAL OF VIROLOGICAL METHODS

Elsevier

Año: 1996 vol. 56 p. 19 - 26

0166-0934

Tomato spotted wilt is a serious disease that affects several economically important crops. From the epidemiological point of view and for the development of a successful plan for transgenic resistance plants, the four known Tospouirus species must be discriminated at the molecular level. A RT-PCR assay using primers complementary to the N gene was used to detect and differentiate fourteen Argentinian isolates of Tospovirus from different crops and geographical areas. Extracts were reverse transcribed using a thermo-resistant reverse transcriptase and PCR reactions were performed for 30 min in a capillar thermo-cycler. The products were digested with restriction enzymes and three of the four described species were identified. Additionaly, the results were confirmed by DAS-ELISA. The method described here is rapid and reliable.Tospouirus species must be discriminated at the molecular level. A RT-PCR assay using primers complementary to the N gene was used to detect and differentiate fourteen Argentinian isolates of Tospovirus from different crops and geographical areas. Extracts were reverse transcribed using a thermo-resistant reverse transcriptase and PCR reactions were performed for 30 min in a capillar thermo-cycler. The products were digested with restriction enzymes and three of the four described species were identified. Additionaly, the results were confirmed by DAS-ELISA. The method described here is rapid and reliable.