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Genotypic Inference of HIV-1 Tropism Using Population-based Sequencing of V3
Published on: December 27, 2010
Development and Validation of a New Set of Primers for Identification of Circulating Lineages and
María Paula Reyes-Zuluaga1, José Antonio Pérez-Pérez1,2, Wilson Correia1
1Instituto Universitario de Enfermedades Tropicales y Salud Pública de Canarias, Universidad de La Laguna, 38200 La Laguna, Tenerife, Spain.
Insights
Human Respiratory Syncytial Virus (HRSV) is a major cause of respiratory illness in young children in Cabo Verde. Researchers developed a method to detect HRSV lineages and mutations, crucial for monitoring vaccine effectiveness.
Area of Science:
- Virology
- Public Health
- Molecular Biology
Background:
- Acute Respiratory Infection (ARI) is a leading cause of mortality in children under 5 in Cabo Verde.
- Human Respiratory Syncytial Virus (HRSV) is a primary etiological agent of ARI in this population.
- Limited epidemiological data on HRSV in Cabo Verde hinders public health assessments.
Purpose of the Study:
- To develop and validate a primer set for HRSV G and F gene amplification and sequencing.
- To identify circulating HRSV lineages in Cabo Verde.
- To detect mutations in the HRSV F protein relevant to prophylactic therapies.
Main Methods:
- Development and validation of a primer collection for HRSV G and F genes.
- Utilized conventional and semi-nested PCR for amplification.
- Employed Sanger sequencing for genetic analysis.
Main Results:
- Successfully identified HRSV lineages A.D.2.2.1, A.D.3, B.D.4.1.1, and B.D.E.1.
- Detected globally circulating HRSV lineages during 2019 and 2022.
- Found no mutations conferring resistance to Palivizumab or Nirsevimab.
Conclusions:
- The developed method enables HRSV lineage and mutation detection in Cabo Verde.
- Continuous HRSV genotype monitoring is essential for public health.
- Ensuring the efficacy of prophylactic therapies like Palivizumab and Nirsevimab requires ongoing surveillance.
Abstract:
In Cabo Verde, Acute Respiratory Infection caused by various pathogens was the most reported condition in children under 5 years old between 2014-2020, and the fourth leading cause of mortality in this age group, with Human Respiratory Syncytial Virus (HRSV) being one of the main etiological agents. However, limited literature on the subject hinders the study of its epidemiology and the evaluation of potential implications for public health. In this work, we developed and validated a primer collection for the amplification and sequencing of the G and F genes of HRSV, using a sequential workflow including conventional and semi-nested PCR, followed by Sanger sequencing. This strategy not only allowed for the identification of HRSV linages but also facilitated the detection of mutants in the HRSV F protein, a critical step towards evaluating and ensuring the continued efficacy of Nirsevimab or Palivizumab as prophylactic therapies. Our analysis revealed the presence of the HRSV lineages A.D.2.2.1, A.D.3, B.D.4.1.1, and B.D.E.1, corresponding to the globally circulating lineages during the study period (years 2019 and 2022). No previously described mutations in the F protein that confer resistance to Palivizumab and Nirsevimab were found. However, continuous monitoring of HRSV genotypes is crucial to promptly identifying resistant viruses, considering their potential impact on public health.
