Phylogenetic and evolutionary analysis of dengue virus serotypes circulating at the Colombian-Venezuelan border during 2015-2016 and 2018-2019.

Dengue is an endemic disease in Colombia. Norte de Santander is a region on the border of Colombia and Venezuela and has reported the co-circulation and simultaneous co-infection of different serotypes of the dengue virus (DENV). This study aimed to conduct a phylogenetic analysis on the origin and genetic diversity of DENV strains circulating in this bordering region. Serum samples were collected from patients who were clinically diagnosed with febrile syndrome associated with dengue during two periods. These samples were tested for DENV and serotyping was performed using reverse transcriptase-polymerase chain reaction. Subsequently, positive samples were amplified and the envelope protein gene of DENV was sequenced. Phylogenetic and phylogeographic analyses were performed using the sequences obtained. Basic local alignment search tool analysis confirmed that six and eight sequences belonged to DENV-1 and DENV-2, respectively. The phylogenetic analysis of DENV-1 showed that the sequences belonged to genotype V and clade I; they formed two groups: in the first group, two sequences showed a close phylogenetic relationship with strains from Ecuador and Panama, whereas the other four sequences were grouped with strains from Venezuela and Colombia. In the case of DENV-2, the analysis revealed that the sequences belonged to the Asian-American genotype and clade III. Furthermore, they formed two groups; in the first group, three sequences were grouped with strains from Colombia and Venezuela, whereas the other five were grouped with strains from Venezuela, Colombia and Honduras. This phylogenetic analysis suggests that the geographical proximity between Colombia and Venezuela is favourable for the export and import of different strains among serotypes or clades of the same DENV serotype, which could favour the spread of new outbreaks caused by new strains or genetic variants of this arbovirus. Therefore, this information highlights the importance of monitoring the transmission of DENV at border regions.

Co-circulation and simultaneous co-infection of dengue, chikungunya, and zika viruses in patients with febrile syndrome at the Colombian-Venezuelan border.

BACKGROUND: In Colombia, the dengue virus (DENV) has been endemic for decades, and with the recent entry of the chikungunya virus (CHIKV) (2014) and the Zika virus (ZIKV) (2015), health systems are overloaded because the diagnosis of these three diseases is based on clinical symptoms, and the three diseases share a symptomatology of febrile syndrome. Thus, the objective of this study was to use molecular methods to identify their co-circulation as well as the prevalence of co-infections, in a cohort of patients at the Colombian-Venezuelan border. METHODS: A total of 157 serum samples from patients with febrile syndrome consistent with DENV were collected after informed consent and processed for the identification of DENV (conventional PCR and real-time PCR), CHIKV (conventional PCR), and ZIKV (real-time PCR). DENV-positive samples were serotyped, and some of those positive for DENV and CHIKV were sequenced. RESULTS: Eighty-two patients were positive for one or more viruses: 33 (21.02%) for DENV, 47 (29.94%) for CHIKV, and 29 (18.47%) for ZIKV. The mean age range of the infected population was statistically higher in the patients infected with ZIKV (29.72 years) than in those infected with DENV or CHIKV (21.09 years). Both co-circulation and co-infection of these three viruses was found. The prevalence of DENV/CHIKV, DENV/ZIKV, and CHIKV/ZIKV co-infection was 7.64%, 6.37%, and 5.10%, with attack rates of 14.90, 12.42, and 9.93 cases per 100,000 inhabitants, respectively. Furthermore, three patients were found to be co-infected with all three viruses (prevalence of 1.91%), with an attack rate of 4.96 cases per 100,000 inhabitants. CONCLUSION: Our results demonstrate the simultaneous co-circulation of DENV, CHIKV, ZIKV and their co-infections at the Colombian-Venezuelan border. Moreover, it is necessary to improve the differential diagnosis in patients with acute febrile syndrome and to study the possible consequences of this epidemiological overview of the clinical outcomes of these diseases in endemic regions.