Real-time RT-PCR for Venezuelan equine encephalitis complex, Madariaga, and Eastern equine encephalitis viruses: application in human and mosquito public health surveillance in Panama.

Eastern equine encephalitis virus (EEEV), Madariaga virus (MADV), and Venezuelan equine encephalitis virus complex (VEEV) are New World alphaviruses transmitted by mosquitoes. They cause febrile and sometimes severe neurological diseases in human and equine hosts. Detecting them during the acute phase is hindered by non-specific symptoms and limited diagnostic tools. We designed and clinically assessed real-time reverse transcription polymerase chain reaction assays (rRT-PCRs) for VEEV complex, MADV, and EEEV using whole-genome sequences. Validation involved 15 retrospective serum samples from 2015 to 2017 outbreaks, 150 mosquito pools from 2015, and 118 prospective samples from 2021 to 2022 surveillance in Panama. The rRT-PCRs detected VEEV complex RNA in 10 samples (66.7%) from outbreaks, with one having both VEEV complex and MADV RNAs. VEEV complex RNA was found in five suspected dengue cases from disease surveillance. The rRT-PCR assays identified VEEV complex RNA in three Culex (Melanoconion) vomerifer pools, leading to VEEV isolates in two. Phylogenetic analysis revealed the VEEV ID subtype in positive samples. Notably, 11.9% of dengue-like disease patients showed VEEV infections. Together, our rRT-PCR validation in human and mosquito samples suggests that this method can be incorporated into mosquito and human encephalitic alphavirus surveillance programs in endemic regions.

SARS-CoV-2 reinfection with a virus harboring mutation in the Spike and the Nucleocapsid proteins in Panama.

We report a case of reinfection by SARS-CoV-2 with the second virus harboring amino acid changes in the Spike protein (141-143del, D215A, ins215AGY, L452R, D614G), orf1a, helicase, orf3a, and Nucleocapside. The virus associated with the reinfection, from an endemic lineage containing the S:L452R immune escape mutation, was circulating in Panama at the time.

Severe acute respiratory syndrome coronavirus 2 detected in placentas of 2 coronavirus disease 2019-positive asymptomatic pregnant women-case report.

There is limited evidence regarding severe acute respiratory syndrome coronavirus 2 infection in the placenta of pregnant women who tested positive, and if this could be a route for vertical transmission of the virus in utero. We present the cases of 2 pregnant women in their third trimester who were admitted for delivery by cesarean delivery and who, through universal screening, tested positive for coronavirus disease 2019. The maternal and fetal sides of the placenta were sectioned from both patients for viral analysis. Real-time polymerase chain reaction analysis of the placental-extracted RNA revealed a severe acute respiratory syndrome coronavirus 2 infection on the fetal side of the placenta in both patients. The virus was isolated from the patient with the lowest cycle threshold value on the fetal side of the placenta. Whole genome sequencing showed that the virus detected in this placenta was from the B1 lineage. Immunohistochemical analysis of the placental tissue detected severe acute respiratory syndrome coronavirus 2 in the endothelial cells of chorionic villi vessels proximal to both the maternal and fetal sides, with a granular cytoplasmic pattern and perinuclear reinforcement. Histologic examination of the placenta also detected a dense infiltrate of lymphoid cells around decidual vessels and endothelial cells with cytopathic changes, especially on the maternal side. Nasopharyngeal swabs from the infants that were subjected to reverse transcription quantitative polymerase chain reaction testing were negative for severe acute respiratory syndrome coronavirus 2 at 24 hours after birth. A follow-up analysis of the infants for immunoglobin G and immunoglobin M expression, clinical manifestations, and long-term developmental abnormalities is recommended.

Molecular Epidemiology of Dengue in Panama: 25 Years of Circulation.

Dengue virus (DENV) is the most prevalent arbovirus in terms of human public health importance globally. In addition to DENV epidemiological surveillance, genomic surveillance may help investigators understand the epidemiological dynamics, geographic distribution, and temporal patterns of DENV circulation. Herein, we aimed to reconstruct the molecular epidemiology and phylogeny of DENV in Panama to connect the epidemiological history of DENV dispersal and circulation in Latin America. We retrospectively analyzed the epidemiological data obtained during 25 years of DENV surveillance in Panama. DENV was reintroduced in Panama in 1993 after a 35 year absence of autochthonous transmission. The increase in the number of total dengue cases has been accompanied by an increase in severe and fatal cases, with the highest case fatality rate recorded in 2011. All four serotypes were detected in Panama, which is characterized by serotype replacement and/or co-circulation of multiple serotypes. Phylogenetic analysis of datasets collected from envelope (E) gene sequences obtained from viruses isolated from human sera demonstrated that circulating viruses were highly diverse and clustered in distinct clades, with co-circulation of clades from the same genotype. Our analyses also suggest that Panamanian strains were related to viruses from different regions of the Americas, suggesting a continuous exchange of viruses within the Americas.