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• W2000950736 abstract "HIV-1 subtype B is the predominant genetic form in western Europe, but non-B subtype and intersubtype recombinant strains are being identified with increasing frequency, mainly in natives of African countries [1]. It is of particular interest to know which are the genetic forms circulating in an area in order to know the potential for spread of any new variant, as has happened in Thailand [2], Kaliningrad [3] or Argentina [4]. Previous studies in Spain have described the presence of non-B subtype HIV-1 strains in inmigrants [5] and in natives [6]. Linked with the last study, we have reported the prevalence of drug resistance mutations in B, non-B subtypes and recombinant forms of HIV-1 in infected individuals in Spain [7]. In order to describe the patterns of reverse transcriptase (RT) and protease-associated mutations in non-B subtypes and recombinant forms of HIV-1, we analysed all the different RT and protease amino acid substitutions in HIV-1 of the Spanish patients previously reported [6,7], together with 115 newly identified viruses. In total, 556 HIV-1-infected individuals were included. RNA extraction from plasma was performed by the Boom method and RT and protease genes were amplified and sequenced as reported [8]. Subtype sequence homologies were examined using the BLAST algorithm with ‘Genotyping Tool’ (http://www. ncbi.nln.gov/retroviruses/subtype/subtype.htlm). Non-B subtype sequences were further examined by neighbouring phylogenetic trees, and intersubtype recombination was analysed by bootscanning using Simplot. Primary and secondary resistance mutations were defined following the criteria reported [9,10]. The statistical analysis was performed by McNemar test (RSIGMA, Babel statistical computer programme). Non-B subtype and recombinant viruses were identified in 31 (5.6%) of the 556 samples analysed, 20 of these had been reported previously [6] and 11 correspond to newly identified viruses. In one specific province (Pontevedra), the prevalence was 8.9% most probably because of its large fishing port with intensive contact with African ports, mainly in the Guinea Gulf, and close contact with Portugal. It is important to stress that 25 patients were native Spanish (80.6%), three were Portuguese, two were African and one was Argentinian. Seventeen (54.8%) were injecting drug users (IDU) (nine also had heterosexual exposure), and 14 were infected by heterosexual contact. Seven men had had sexual contacts with African women, three women were prostitutes, and five men were sailors who had multiple heterosexual contacts, but 10 (32%) were Spanish without any contact with Africans. The different non-B and recombinant HIV-1 viruses detected were: G, 11 (36%); A, two (6%); C, one (3%); H, one (3%); BG, four (13%); dual B + G infection, one (3%); UG, four (13%); GH, one (3%); UK, one (3%); DF, one (3%); BF, one (3%); and UAJ, three (10%). The proportion of primary resistance mutations was similar among B and non-B samples, as we have previously described [7], and as documented in other studies [11–13], in relation with some resistance mutations. Twenty-one (67%) of these 31 non-B strains were G subtype or intersubtype recombinants with segments of subtype G. Nineteen (90.5%) were IDU. Fifteen (71.4%) were from the province of Pontevedra. An epidemiological link between these 21 individuals was known in only two patients (X-138 and X-279) who were sexual partners. A possible initial source of these G subtype viruses in Galicia could be through sexual contact of seamen with African women in African seaports, with a subsequent spread in Spain among IDU and their sexual partners. All 12 subtype G sequences (including G subtype clones of a dually infected individual) and four BG recombinant intersubtypes formed a monophyletic cluster supported by a high (93%) bootstrap value (when B subtype regions of the recombinants were excluded), indicating a common ancestry. The clinical and epidemiological data of these 16 patients are shown in Table 1. Thirteen (81.2%) were Spanish, and IDU was the risk exposure associated with HIV-1 infection. Twelve (75%) patients were treatment-naive, and four were under highly active antiretroviral therapy.Table 1: Epidemiological, clinical and virological data on HIV-1-infected patients with subtype G and GB recombinants in Spain (Galicia). The analysis of secondary mutations and polymorphisms showed that all subtype G viruses of the monophyletic group and one G subtype clone of the dually infected individual had the same RT and protease mutations (Table 1). Statistically significant differences were found in the frequency of these mutations between B and G subtype viruses (P < 0.001), being higher among G than among B subtype viruses. Two RT substitutions, A98S and R211S, were specifically detected in the viruses of the subtype G monophyletic group, but in none of the other non-B subtype or recombinant viruses included in the study. To our knowledge, none of these two mutations have been reported as characteristic of other subtype G strains, suggesting that they could be a signature of subtype G viruses circulating in this area of Spain. Two protease mutations, M46L, associated with resistance to indinavir [14], and 19P, were detected in three subtype G viruses and one G subtype clone of the dually infected individual. These four viruses were isolated from treatment-naive patients. To our knowledge, this is the first time these mutations have been detected in viruses from non-treated patients. The K20I substitution in protease has been published as a signature of subtype G [15]. We detected this mutation in subtype G but also in other non-B subtype viruses. To determine the clinical relevance of these findings, phenotypic resistance testing and biological characterization of G subtype viruses are now in progress. These data could reveal differences between B and G subtypes, and their impact on the use of antiretroviral drugs, allowing for a more reliable interpretation of resistance assays in the treatment of patients infected with non-B subtypes. Acknowledgements The authors would like to thank Dr José María Hernández Cochón, Conselleiro de Sanidade e Servicios Sociais and Dra Pilar Farjas Abadía Directora Xeral de Saúde Pública, Xunta de Galicia for their support in the development of the study in Galicia. The technical assistance of Isabel Herrero, Milagros Pinilla and Valentina García is gratefully acknowledged. *The Spanish Group for Antiretroviral Resistance Studies in Galicia: A. Agulla, A. Mariño, Hospital Arquitecto Marcide, Ferrol (La Coruña); S. López-Calvo, J.D. Pedreira, Hospital Juan Canalejo (La Coruña); A. Aguilera, E. Losada, A. Prieto, Complejo Hospitalario Universitario de Santiago (La Coruña); J. Corredoira, M.J. López-Alvarez, A. Rodríguez, Hospital Xeral-Calde (Lugo); J. García-Costa, R. Fernández-Rodríguez, Hospital Nuestra Señora del Cristal, (Orense); R. Rodríguez, Hospital Provincial Santa María Madre (Orense); C. Miralles, A. Ocampo, Hospital Xeral-Cíes, Vigo (Pontevedra); R. Ojea de Castro, Hospital Montecelo (Pontevedra); L.E. Morano, R. Pérez-Rodríguez, A. Rodríguez, J. Torres, Hospital Meixoeiro, Vigo (Pontevedra); J. Díz, R. Rodríguez-Real, Hospital Xeral Provincial (Pontevedra). Lucía Pérez-Alvareza Michael M. Thomsona María Luisa Villahermosaa Elena Vázquez de Pargaa Aurora Rodríguezb María Teresa Cuevasa Elena Delgadoa Nuria Manjóna Celia Mirallesb Leandro Medranoa Jose Antonio Taboadab Rafael Nájeraa and the Spanish Group for Antiretroviral Resistance Studies in Galicia*" @default.
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• W2000950736 title "HIV-1 subtype G and BG recombinant viruses in Spanish natives: evidence of characteristic mutations in reverse transcriptase and protease" @default.
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