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      Rapid Multiplexed Immunoassay for Detection of Antibodies to Kaposi’s Sarcoma-Associated Herpesvirus

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          Abstract

          Diagnosis of KSHV-infected individuals remains a challenge. KSHV prevalence is high in several populations with high prevalence of HIV, leading to increased risk of development of Kaposi’s sarcoma (KS). While current assays are reliable for detecting antibodies to KSHV, none are routinely utilized to identify individuals with KSHV infection and thus at increased risk for KS due to assay complexity, lack of access to testing, and cost, particularly in resource-limited settings. Here we describe the addition of KSHV proteins LANA and K8.1 to a previously evaluated HIV/co-infection multiplexed fluorescence immunoassay system. This study demonstrates assay performance by measuring antibody reactivity for KSHV and HIV-1 in a collection of clinical specimens from patients with biopsy-proven KS and sourced negative controls. The KSHV assay correctly identified 155 of 164 plasma samples from patients with biopsy-proven KS and 85 of 93 KSHV antibody (Ab)-negative samples for a sensitivity of 95.1% and specificity of 91.4%. Assay performance for HIV-1 detection was also assessed with 100% agreement with independently verified HIV-1 Ab-positive and Ab-negative samples. These results demonstrate good sensitivity and specificity for detection of antibody to KSHV antigens, and demonstrate the potential for multiplexed co-infection testing in resource-limited settings to identify those at increased risk for HIV-1-related complications.

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          Identification of herpesvirus-like DNA sequences in AIDS-associated Kaposi's sarcoma.

          Representational difference analysis was used to isolate unique sequences present in more than 90 percent of Kaposi's sarcoma (KS) tissues obtained from patients with acquired immunodeficiency syndrome (AIDS). These sequences were not present in tissue DNA from non-AIDS patients, but were present in 15 percent of non-KS tissue DNA samples from AIDS patients. The sequences are homologous to, but distinct from, capsid and tegument protein genes of the Gammaherpesvirinae, herpesvirus saimiri and Epstein-Barr virus. These KS-associated herpesvirus-like (KSHV) sequences appear to define a new human herpesvirus.
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            KSHV infection and the pathogenesis of Kaposi's sarcoma.

            Don Ganem (2006)
            Kaposi's sarcoma (KS) has long been suspected of having an infectious etiology on the basis of its unusual epidemiology, histopathology, and natural history. Nearly a decade ago, a novel herpesviral genome was discovered in KS biopsies, and since that time strong epidemiologic evidence has accumulated correlating infection with this KS-associated herpesvirus (KSHV, also known as human herpesvirus 8) with the development of the disease. Here we review the evidence linking KSHV infection to KS risk and discuss current notions of how KSHV gene expression promotes the development of this remarkable neoplasm. These studies show that both latent and lytic viral replicative cycles contribute significantly-but differently-to KS development. The studies also highlight mechanistic differences between oncogenesis caused by KSHV and that caused by its distant relative Epstein-Barr virus.
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              Trends in the incidence of cancer in the black population of Harare, Zimbabwe 1991-2010.

              Incidence rates of different cancers have been calculated for the black population of Harare, Zimbabwe for a 20-year period (1991-2010) coinciding with continuing social and lifestyle changes, and the peak, and subsequent wane, of the HIV-AIDS epidemic. The overall risk of cancer increased during the period in both sexes, with rates of cervix and prostate cancers showing particularly dramatic increases (3.3% and 6.4% annually, respectively). By 2004, prostate cancer had become the most common cancer of men. The incidence of cancer of the esophagus, formerly the most common cancer of men, has remained relatively constant, whereas rates of breast and cervix cancers, the most common malignancies of women, have shown significant increases (4.9% and 3.3% annually, respectively). The incidence of Kaposi sarcoma increased to a maximum around 1998-2000 and then declined in all age groups, and in both sexes The incidence of squamous cell cancers of the conjunctiva is relatively high, with temporal trends similar to those of Kaposi sarcoma. Non-Hodgkin lymphoma, the fifth most common cancer of men and fourth of women, showed a steady increase in incidence throughout the period (6.7-6.9% annually), although rates in young adults (15-39) have decreased since 2001. Cancer control in Zimbabwe, as elsewhere in sub-Saharan Africa, involves meeting the challenge of emerging cancers associated with westernization of lifestyles (large bowel, breast and prostate), while the incidence of cancers associated with poverty and infection (liver, cervix and esophagus) shows little decline, and the residual burden of the AIDS-associated cancers remains significant. Copyright © 2013 UICC.
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                Author and article information

                Contributors
                Role: Editor
                Journal
                PLoS One
                PLoS ONE
                plos
                plosone
                PLoS ONE
                Public Library of Science (San Francisco, CA USA )
                1932-6203
                26 September 2016
                2016
                : 11
                : 9
                : e0163616
                Affiliations
                [1 ]Department of Medicine, Division of Infectious Diseases, University of California San Diego, San Diego, CA, United States of America
                [2 ]MBio Diagnostics, Inc., Boulder, CO, United States of America
                [3 ]Department of Medicine, Division of Infectious Diseases, University of Colorado Denver, Aurora, CO, United States of America
                [4 ]Department of Immunology and Microbial Science, The Scripps Research Institute, La Jolla, CA, United States of America
                [5 ]Department of Medicine, University of Zimbabwe College of Health Sciences, Harare, Zimbabwe
                [6 ]Departments of Medicine and Medical and Laboratory Sciences, University of Zimbabwe College of Health Sciences, Harare, Zimbabwe
                Institut Pasteur of Shanghai Chinese Academy of Sciences, CHINA
                Author notes

                Competing Interests: This manuscript is a result of the collaboration between authors from the University of California, San Diego, the University of Colorado, and the commercial organization MBio Diagnostics, Inc. Funding was provided by the NIH directly to MBio Diagnostics under a Small Business Innovative Research grant for development of diagnostic technologies. Two authors of the manuscript are current employees of MBio Diagnostics, Inc. The authors’ affiliation does not alter our adherence to PLOS ONE policies on sharing data and materials.

                • Conceived and designed the experiments: MJL CAB RTS.

                • Performed the experiments: CL KT.

                • Analyzed the data: CL KT SPF TBC.

                • Contributed reagents/materials/analysis tools: TBC MB IG LG BN JHE.

                • Wrote the paper: CL JHE RTS.

                Article
                PONE-D-16-04637
                10.1371/journal.pone.0163616
                5036886
                27669509
                cfc4faaa-8ef1-46e8-8df5-748d7cc33ad7
                © 2016 Logan et al

                This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

                History
                : 2 February 2016
                : 12 September 2016
                Page count
                Figures: 2, Tables: 2, Pages: 13
                Funding
                Funded by: funder-id http://dx.doi.org/10.13039/100000060, National Institute of Allergy and Infectious Diseases;
                Award ID: R44AI068543
                Award Recipient :
                Research reported in this publication was supported by the National Institute of Allergy and Infectious Diseases of the National Institutes of Health under award number R44AI068543, provided directly to MBio Diagnostics, Inc. The funders provided support in the form of salaries for authors KT and MJL, but did not have any additional role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. The specific roles of these authors are articulated in the ‘author contributions’ section. The content of this manuscript is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health.
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                Research and Analysis Methods
                Immunologic Techniques
                Immunoassays
                Enzyme-Linked Immunoassays
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                Kaposi's Sarcoma-Associated Herpesvirus
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