Thrips-transmitted tospoviruses (Tospovirus: Bunyaviridae) cause serious losses in numerous agronomic and horticultural crops. In November 2015, tomato and bell pepper plants with foliar symptoms consisting of necrotic lesions, necrotic rings, veinal necrosis, and stem necrosis were collected in grower fields in the states of Antioquia and Risaralda (0 to 40% of incidence). Symptomatic leaves of three tomato plants and fruits of two bell peppers from Antioquia, and leaves of six tomato plants from Risaralda were first tested for the presence of TSWV by DAS-ELISA (Agdia Inc., Elkhart, IN). Additionally, six tomato plants from Risaralda were tested by TSWV-specific immunostrips (Agdia Inc.). The two bell peppers and one tomato plant from Antioquia were positive by ELISA, while four tomato plants from Risaralda were positive by immunostrip and two of those by ELISA. ELISA and/or Immunostrip-positive samples were further tested by RT-PCR with primers specific to the TSWV nucleoprotein (Bag et al. 2012). However, no amplicons were obtained. A degenerate primer pair specific to the polymerase gene of tospoviruses (Chu et al. 2001) was used. Amplicons of expected size (∼900 bp) were obtained from all the ELISA positive samples. Four of them were cloned, and three independent clones per amplicon were sequenced in both directions (GenBank accession nos. KX833214, KX833215, and KX833216). GenBank search showed nucleotide sequence identity of 83% with TSWV (D10066), 81% with Groundnut ring spot virus (KT972590) and Tomato chlorotic spot virus (HQ700667.1), and 75% with Impatiens necrotic spot virus (X93218.1). To identify the Colombian tospovirus to the species level, a set of generic primers (Hassani-Mehraban et al. 2016) that amplify a ∼750 bp region of the small RNA (partial N gene and its 3′ UTR region) of tospoviruses belonging to the American clade was used. Three out of four samples produced the amplicon of expected size, and one was cloned and sequenced. The consensus sequence (KX833218) showed an identity of 99% with the corresponding region of the N gene of Alstroemeria necrotic streak virus (ANSV; GQ478668). ANSV was previously reported infecting alstroemeria plants from Colombia (Hassani-Mehraban et al. 2010). To our knowledge, this is the first report of natural infection of tomato and bell pepper plants by ANSV in Colombia, and the first report of the partial sequence of the L gene of ANSV. TSWV antiserum cross reacts with ANSV, GRSV, and TCSV (Hassani-Mehraban et al. 2016), and the immunostrips cross react with GRSV and TCSV (Gilbertson et al. 2015). This was the reason why samples were positive for TSWV by serology but negative by RT-PCR. The limitations of the methods (no specific antiserum, limited sequence information) available to detect ANSV underscore the need to use different sets of primers followed by sequence comparisons for accurate identification of tospoviruses (Gilbertson et al. 2015). The expanding host range of ANSV could raise the phytosanitary risk for solanaceous and ornamental crops in Colombia, and highlights the need for continuous surveys and the use of species-specific detection tools in order to formulate accurate virus management tactics.