Viremia Associated with Fatal Outcomes in Ferrets Infected with Avian H5N1 Influenza Virus

Avian H5N1 influenza viruses cause severe disease and high mortality in infected humans. However, tissue tropism and underlying pathogenesis of H5N1 virus infection in humans needs further investigation. The objective of this work was to study viremia, tissue tropism and disease pathogenesis of H5N1 virus infection in the susceptible ferret animal model. To evaluate the relationship of morbidity and mortality with virus loads, we performed studies in ferrets infected with the H5N1 strain A/VN/1203/04 to assess clinical signs after infection and virus load in lung, brain, ileum, nasal turbinate, nasal wash, and blood. We observed that H5N1 infection in ferrets is characterized by high virus load in the brain and and low levels in the ileum using real-time PCR. In addition, viral RNA was frequently detected in blood one or two days before death and associated with symptoms of diarrhea. Our observations further substantiate pathogenicity of H5N1 and further indicate that viremia may be a bio-marker for fatal outcomes in H5N1 infection

Analysis of Healthcare Workers' Knowledge About Vital Sign Zero and Identify-Isolate-Inform (3I) System in the Diagnosis and Prevention of Infectious Diseases in Chinese Tertiary Hospitals

Objective To explore the current knowledge and application of vital sign zero and the identify-isolate-inform (3I) system among healthcare workers in China in order to provide a reference for future improvement of healthcare workers' awareness of personal protection and prevention and control measures of infectious diseases. Methods The questionnaire was used to investigate the basic information of health care workers, their knowledge and application of Vital sign zero and the 3I system. A total of 602 forms of health care workers from tertiary hospitals were randomly collected and included for analysis. Results The survey showed that 45.30% and 57.30% of the healthcare workers from Chinese tertiary hospitals know about vital sign zero and 3I system while 51.80% and 57.30% of them applied these measures in their clinical practices. Logistics regression analysis results showed that healthcare workers aged 35 years old and below were less aware of vital sign zero than those above 50 years old (OR = 0.405, 95% CI: 0.174–0.942, P = 0.036). Compared with those in Northwest China, healthcare workers who worked in East China (OR = 0.147, 95% CI: 0.031–0.702, P = 0.016), Central China (OR = 0.085, 95% CI: 0.018–0.403, P = 0.002), Southwest China (OR = 0.083, 95% CI: 0.014–0.48, P = 0.006) and North China (OR = 0.201, 95% CI: 0.042–0.966, P = 0.045) were less aware of vital sign zero while the healthcare workers in Northeast China (OR=9.714, 95% CI: 1.091–86.521, P = 0.042), East China (OR = 18.049, 95% CI: 2.258–144.259, P = 0.006), Central China (OR = 25.560, 95% CI: 3.210–203.502, P = 0.002), South China (OR = 11.141, 95% CI: 1.395–88.947, P = 0.023), Southwest China (OR = 23.200, 95% CI: 2.524–213.286, P = 0.005) and North China (OR = 14.078, 95% CI: 1.756–112.895, P = 0.013) had a better understanding of the 3I system than those in Northwest China. Healthcare workers with more than 20 years of working experience showed less knowledge of the 3I system than those with less than 5 years of working experience (OR = 0.409, 95% CI: 0.215–0.77, P = 0.006). Conclusion The current levels of knowledge and application of vital sign zero and the 3I system in the healthcare workers of Chinese tertiary hospitals need to be improved. The concept of vital sign zero should be incorporated into the prevention triage system of infectious diseases

Identification of new, emerging HIV-1 unique recombinant forms and drug resistant viruses circulating in Cameroon

<p>Abstract</p> <p>Background</p> <p>The HIV epidemic in Cameroon is characterized by a high degree of viral genetic diversity with circulating recombinant forms (CRFs) being predominant. The goal of our study was to determine recent trends in virus evolution and emergence of drug resistance in blood donors and HIV positive patients.</p> <p>Methodology</p> <p>Blood specimens of 73 individuals were collected from three cities and a few villages in Cameroon and viruses were isolated by co-cultivation with PBMCs. Nested PCR was performed for gag p17 (670 bp) pol (840 bp) and Env gp41 (461 bp) genes. Sequences were phylogenetically analyzed using a reference set of sequences from the Los Alamos database.</p> <p>Results</p> <p>Phylogenetic analysis based on partial sequences revealed that 65% (n = 48) of strains were CRF02_AG, 4% (n = 3) subtype F2, 1% each belonged to CRF06 (n = 1), CRF11 (n = 1), subtype G (n = 1), subtype D (n = 1), CRF22_01A1 (n = 1), and 26% (n = 18) were Unique Recombinant Forms (URFs). Most URFs contained CRF02_AG in one or two HIV gene fragments analyzed. Furthermore, pol sequences of 61 viruses revealed drug resistance in 55.5% of patients on therapy and 44% of drug naïve individuals in the RT and protease regions. Overall URFs that had a primary HIV subtype designation in the pol region showed higher HIV-1 p24 levels than other recombinant forms in cell culture based replication kinetics studies.</p> <p>Conclusions</p> <p>Our results indicate that although CRF02_AG continues to be the predominant strain in Cameroon, phylogenetically the HIV epidemic is continuing to evolve as multiple recombinants of CRF02_AG and URFs were identified in the individuals studied. CRF02_AG recombinants that contained the pol region of a primary subtype showed higher replicative advantage than other variants. Identification of drug resistant strains in drug-naïve patients suggests that these viruses are being transmitted in the population studied. Our findings support the need for continued molecular surveillance in this region of West Central Africa and investigating impact of variants on diagnostics, viral load and drug resistance assays on an ongoing basis.</p

SARS-CoV-2 incidence, seroprevalence, and COVID-19 vaccination coverage in the homeless population: a systematic review and meta-analysis

ObjectivesSARS-CoV-2 infection and COVID-19 vaccination of homeless people are a serious public health concern during COVID-19 pandemic. We aimed to systematically assess SARS-CoV-2 incidence, seroprevalence, and COVID-19 vaccination coverage in homeless people, which are important to inform resource allocation and policy adjustment for the prevention and control of COVID-19.MethodsWe searched PubMed, Web of Science, and the World Health Organization COVID-19 database for the studies of SARS-CoV-2 incidence, seroprevalence, and COVID-19 vaccination coverage in the homeless population. Subgroup analyses were conducted to pool SARS-CoV-2 incidence and seroprevalence in sheltered homeless, unsheltered homeless, and mixed population, respectively. Potential sources of heterogeneity in the estimates were explored by meta-regression analysis.ResultsForty-nine eligible studies with a total of 75,402 homeless individuals and 5,000 shelter staff were included in the meta-analysis. The pooled incidence of SARS-CoV-2 infection was 10% (95% CI: 7 to 12%) in the homeless population and 8% (5 to 12%) for shelter staff. In addition, the overall estimated SARS-CoV-2 specific seroprevalence was 19% (8 to 33%) for homeless populations and 22% (3 to 52%) for shelter staff, respectively. Moreover, for the homeless subjects, the pooled incidence was 10% (4 to 23%) for asymptomatic SARS-CoV-2 infections, 6% (1 to 12%) for symptomatic SARS-CoV-2 infections, 3% (1 to 4%) for hospitalization for COVID-19, and 1% (0 to 2%) for severe COVID-19 cases, respectively while no COVID-19-related death was reported. Furthermore, the data derived from 12 included studies involving 225,448 homeless individuals revealed that the pooled proportion of one dose COVID-19 vaccination was 41% (35 to 47%), which was significantly lower than those in the general population.ConclusionOur study results indicate that the homeless people remain highly susceptible to SARS-CoV-2 infection, but COVID-19 vaccination coverage was lower than the general population, underscoring the need for prioritizing vaccine deployment and implementing enhanced preventive measures targeting this vulnerable group

Multiplexed, rapid detection of H5N1 using a PCR-free nanoparticle-based genomic microarray assay

<p>Abstract</p> <p>Background</p> <p>For more than a decade there has been increasing interest in the use of nanotechnology and microarray platforms for diagnostic applications. In this report, we describe a rapid and simple gold nanoparticle (NP)-based genomic microarray assay for specific identification of avian influenza virus H5N1 and its discrimination from other major influenza A virus strains (H1N1, H3N2).</p> <p>Results</p> <p>Capture and intermediate oligonucleotides were designed based on the consensus sequences of the matrix (M) gene of H1N1, H3N2 and H5N1 viruses, and sequences specific for the hemaglutinin (HA) and neuraminidase (NA) genes of the H5N1 virus. Viral RNA was detected within 2.5 hours using capture-target-intermediate oligonucleotide hybridization and gold NP-mediated silver staining in the absence of RNA fragmentation, target amplification, and enzymatic reactions. The lower limit of detection (LOD) of the assay was less than 100 fM for purified PCR fragments and 10³TCID₅₀units for H5N1 viral RNA.</p> <p>Conclusions</p> <p>The NP-based microarray assay was able to detect and distinguish H5N1 sequences from those of major influenza A viruses (H1N1, H3N2). The new method described here may be useful for simultaneous detection and subtyping of major influenza A viruses.</p

Absence of Detectable XMRV and Other MLV-Related Viruses in Healthy Blood Donors in the United States

BACKGROUND: Preliminary studies in chronic fatigue syndrome (CFS) patients and XMRV infected animals demonstrated plasma viremia and infection of blood cells with XMRV, indicating the potential risk for transfusion transmission. XMRV and MLV-related virus gene sequences have also been detected in 4-6% of healthy individuals including blood donors in the U.S. These results imply that millions of persons in the U.S. may be carrying the nucleic acid sequences of XMRV and/or MLV-related viruses, which is a serious public health and blood safety concern. METHODOLOGY/PRINCIPAL FINDINGS: To gain evidence of XMRV or MLV-related virus infection in the U.S. blood donors, 110 plasma samples and 71 PBMC samples from blood donors at the NIH blood bank were screened for XMRV and MLV-related virus infection. We employed highly sensitive assays, including nested PCR and real-time PCR, as well as co-culture of plasma with highly sensitive indicator DERSE cells. Using these assays, none of the samples were positive for XMRV or MLV-related virus. CONCLUSIONS/SIGNIFICANCE: Our results are consistent with those from several other studies, and demonstrate the absence of XMRV or MLV-related viruses in the U.S. blood donors that we studied

XMRV: usage of receptors and potential co-receptors

<p>Abstract</p> <p>Background</p> <p>XMRV is a gammaretrovirus first identified in prostate tissues of Prostate Cancer (PC) patients and later in the blood cells of patients with Chronic Fatigue Syndrome (CFS). Although XMRV is thought to use XPR1 for cell entry, it infects A549 cells that do not express XPR1, suggesting usage of other receptors or co-receptors.</p> <p>Methods</p> <p>To study the usage of different receptors and co- receptors that could play a role in XMRV infection of lymphoid cells and GHOST (GFP- Human osteosarcoma) cells expressing CD4 along with different chemokine receptors including CCR1, CCR2, etc., were infected with XMRV. Culture supernatants and cells were tested for XMRV replication using real time quantitative PCR.</p> <p>Results</p> <p>Infection and replication of XMRV was seen in a variety of GHOST cells, LNCaP, DU145, A549 and Caski cell lines. The levels of XMRV replication varied in different cell lines showing differential replication in different cell lines. However, replication in A549 which lacks XPR1 expression was relatively higher than DU145 but lower than, LNCaP. XMRV replication varied in GHOST cell lines expressing CD4 and each of the co- receptors CCR1-CCR8 and bob. There was significant replication of XMRV in CCR3 and Bonzo although it is much lower when compared to DU145, A549 and LNCaP.</p> <p>Conclusion</p> <p>XMRV replication was observed in GHOST cells that express CD4 and each of the chemokine receptors ranging from CCR1- CCR8 and BOB suggesting that infectivity in hematopoietic cells could be mediated by use of these receptors.</p