Metatranscriptomics Analysis Reveals Diverse Viral RNA in Cutaneous Papillomatous Lesions of Cattle.

Bovine papillomavirus (BPV) is associated with bovine papillomatosis, a disease that forms benign warts in epithelial tissues, as well as malignant lesions. Previous studies have detected a co-infection between BPV and other viruses, making it likely that these co-infections could influence disease progression. Therefore, this study aimed to identify and annotate viral genes in cutaneous papillomatous lesions of cattle. Sequences were obtained from the GEO database, and an RNA-seq computational pipeline was used to analyze 3 libraries from bovine papillomatous lesions. In total, 25 viral families were identified, including Poxviridae, Retroviridae, and Herpesviridae. All libraries shared similarities in the viruses and genes found. The viral genes shared similarities with BPV genes, especially for functions as virion entry pathway, malignant progression by apoptosis suppression and immune system control. Therefore, this study presents relevant data extending the current knowledge regarding the viral microbiome in BPV lesions and how other viruses could affect this disease.

Comparative analysis of different PCR-based strategies for HPV detection and genotyping from cervical samples.

BACKGROUND: Human papillomavirus (HPV) is the main cause of cervical cancer. Polymerase chain reaction (PCR)-based techniques are associated with accurate results with respect to HPV detection and genotyping, being able to identify viral DNA at low levels. However, differences in primer design influence their sensibility and specificity, depending on the HPV type assessed. OBJECTIVE: The aim of the study was to comparatively evaluate the effectiveness of three different PCR-based strategies for HPV detection and genotyping from cervical samples. STUDY DESIGN: The procedures were based on different primer design strategies, using MY09/MY11, EntroA, and type specific multiplex PCR primers. RESULTS: Out of 411 samples of cervical scrapings, 45 (10.9%), 50 (12.2%), and 117 (28.5%) were positive for MY09/MY11, EntroA, and multiplex PCR, respectively. For MY09/MY11 positive samples, 36 were negative for EntroA and 23 for multiplex PCR. For EntroA positive samples, 40 were negative for MY09/MY11 and 26 for multiplex PCR. For multiplex PCR positive samples, 96 were negative for MY09/MY11 and 94 for EntroA. MY09/MY11 identified 12 different HPV types, EntroA detected eight types and multiplex PCR detected 11 HPV types. EntroA primers were able to detect HPV in more samples than MY09/MY11, while multiplex PCR, despite the limited targeted HPV types, presented higher sensibility than the other methods. CONCLUSIONS: The three methods presented different advantages and disadvantages, and the present study reinforces the need to use more than one molecular strategy for HPV detection and genotyping, and the development of novel methods which could overcome the limitations of the existing tests.

Molecular characterization and phylogenetic analysis of Chikungunya virus during the 2016 outbreak in Sergipe, northeastern Brazil.

BACKGROUND: Chikungunya (CHIKV) is an arbovirus transmitted mainly by Aedes aegypti females. CHIKV has been highlighted as the pathogen with the greatest impact due to the high morbidity caused by the infection. In 2016, Brazil experienced an outbreak that affected almost 272 000 people. Here, we performed a molecular characterization and phylogenetic analysis of the CHIKV circulating in 2016 in the state of Sergipe, Brazil. METHODS: A partial region of the E1 gene of 16 CHIKV-positive samples from Sergipe State was amplified and sequenced. RESULTS: All sequences belonged to the East-Central-South-African genotype and three point mutations were verified. Two of them were silent mutations and one was a non-synonymous mutation, which changed lysine to threonine at position 211 in the E1 protein. This mutation was present in 81.2% of the sequences, as well as in other five Brazilian sequences from previous studies. This study found that CHIKV strains circulating in Sergipe during the 2016 outbreak belonged to two different haplotypes. CONCLUSIONS: The strains circulating in Sergipe are phylogenetically close to other Brazilian samples circulating in the northeast and southeast of the country, as well as viruses circulating during the same period in Haiti, indicating the rapid spread of these haplotypes.

Application of an entropy-based computational strategy to identify genomic markers for molecular detection and typing of human papillomavirus.

Human papillomavirus (HPV) is a diverse group of double-stranded DNA viruses that present high tropism for the epithelium and infect keratinocytes. Currently, over 200 viral types have been identified, and almost 40 types preferentially infect the epithelial cells of the genital tract. Infections caused by HPV are the most prevalent viral infections that are sexually transmitted in the world. Given how HPV infection is one of the key factors in the development of cervical cancer, we need to develop more effective diagnostic methods to correctly diagnose patients. The significance of our research is that we have developed and applied a novel computational approach based on entropy to identify phylogenetically informative genomic regions that could be used as markers for the detection and typing of HPV. We have demonstrated that our strategy is capable of finding phylogenetically informative L1 regions to design a primer set that can be used to accurately detect and genotype HPV isolates.

Comparative transcriptomic analysis of bovine papillomatosis.

BACKGROUND: Bovine papillomavirus (BPV) belongs to the Papillomaviridae family and infects epithelial cells of bovines and closely related animals, causing hyperproliferative lesions known as warts or papillomas, which may regress or progress to form benign or malignant tumors. The virus enters the host cell and interacts with it by altering the regulation of genes that are responsible for controlling the cell cycle, thus triggering lesion formation. It is not yet known which host genes are regulated by viral infection. Therefore, the objective of this study was to make use of next-generation RNA sequencing methods to identify differentially expressed genes associated with BPV infection, which might elucidate possible marker genes that could be used to control the disease. RESULTS: Transcriptome analysis revealed that 1343 genes were differentially regulated (FDR < 0.05). A comparison of gene expression in infected and noninfected cows indicated that 655 genes were significantly upregulated, and 688 genes were significantly downregulated. Most differentially expressed genes were associated with BPV infection pathways, which supports the hypothesis that viral infection was the mechanism associated with this regulation. CONCLUSIONS: This is the first study that focused on a large-scale evaluation of gene expression associated with BPV infection, which is important to identify possible metabolic pathways regulated by host genes for lesion development. In addition, novel targets could be identified in order to find ligands that interact with BPV, with the aim of interrupting the infection cycle.