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      Real-time PCR assays for the specific detection of monkeypox virus West African and Congo Basin strain DNA

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          Abstract

          Orthopoxvirus monkeypox (MPXV) forms two distinct clades: the MPXV Congo Basin clade viruses are endemic in the Congo Basin, human illness typically presents with symptoms similar to discrete, ordinary smallpox and has a case fatality rate of approximately 10% in unvaccinated populations; the MPXV West African clade viruses have been isolated in West Africa and appear to cause a less severe, and less inter-human transmissible disease. Recently, monkeypox outbreaks were reported in US and Sudan caused by MPXV West African and Congo Basin strains respectively. These events demonstrated the ability and trend of the virus to exploit new hosts and emerge globally; it also emphasizes the need for the diagnosis of MPXV, especially the ability to distinguish between Congo Basin and West African monkeypox strains. In this study, three new real-time PCR assays based on TaqMan probe technology were reported: the MPXV West African specific, Congo Basin strain specific and MPXV generic assays. The new assays demonstrated good specificity and sensitivity in the validation study with multiple platforms and various PCR reagent kits, and will improve the rapid detection and differentiation of monkeypox infections from other rash illness.

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          The detection of monkeypox in humans in the Western Hemisphere.

          During May and June 2003, an outbreak of febrile illness with vesiculopustular eruptions occurred among persons in the midwestern United States who had had contact with ill pet prairie dogs obtained through a common distributor. Zoonotic transmission of a bacterial or viral pathogen was suspected. We reviewed medical records, conducted interviews and examinations, and collected blood and tissue samples for analysis from 11 patients and one prairie dog. Histopathological and electron-microscopical examinations, microbiologic cultures, and molecular assays were performed to identify the etiologic agent. The initial Wisconsin cases evaluated in this outbreak occurred in five males and six females ranging in age from 3 to 43 years. All patients reported having direct contact with ill prairie dogs before experiencing a febrile illness with skin eruptions. We found immunohistochemical or ultrastructural evidence of poxvirus infection in skin-lesion tissue from four patients. Monkeypox virus was recovered in cell cultures of seven samples from patients and from the prairie dog. The virus was identified by detection of monkeypox-specific DNA sequences in tissues or isolates from six patients and the prairie dog. Epidemiologic investigation suggested that the prairie dogs had been exposed to at least one species of rodent recently imported into the United States from West Africa. Our investigation documents the isolation and identification of monkeypox virus from humans in the Western Hemisphere. Infection of humans was associated with direct contact with ill prairie dogs that were being kept or sold as pets. Copyright 2004 Massachusetts Medical Society
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            A tale of two clades: monkeypox viruses.

            Human monkeypox was first recognized outside Africa in 2003 during an outbreak in the USA that was traced to imported monkeypox virus (MPXV)-infected West African rodents. Unlike the smallpox-like disease described in the Democratic Republic of the Congo (DRC; a Congo Basin country), disease in the USA appeared milder. Here, analyses compared clinical, laboratory and epidemiological features of confirmed human monkeypox case-patients, using data from outbreaks in the USA and the Congo Basin, and the results suggested that human disease pathogenicity was associated with the viral strain. Genomic sequencing of USA, Western and Central African MPXV isolates confirmed the existence of two MPXV clades. A comparison of open reading frames between MPXV clades permitted prediction of viral proteins that could cause the observed differences in human pathogenicity between these two clades. Understanding the molecular pathogenesis and clinical and epidemiological properties of MPXV can improve monkeypox prevention and control.
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              A POX-LIKE DISEASE IN CYNOMOLGUS MONKEYS

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                Author and article information

                Journal
                J Virol Methods
                J Virol Methods
                Journal of Virological Methods
                Elsevier/North-Holland Biomedical Press
                0166-0934
                1879-0984
                17 July 2010
                October 2010
                17 July 2010
                : 169
                : 1
                : 223-227
                Affiliations
                Poxvirus and Rabies Branch, Division of High-Consequence Pathogens and Pathology (Proposed), National Center for Emerging and Zoonotic Infectious Diseases (Proposed), Centers for Disease Control and Prevention, Atlanta, GA, United States
                Author notes
                [* ]Corresponding author at: Poxvirus and Rabies Branch, Division of High-Consequence Pathogens and Pathology (Proposed), National Center for Emerging and Zoonotic Infectious Diseases, Centers for Disease Control and Prevention, Mail Stop G-43, 1600 Clifton Road, NE, Atlanta, GA 30333, United States. Tel.: +1 404 639 2185; fax: +1 404 639 3111.
                Article
                S0166-0934(10)00254-5
                10.1016/j.jviromet.2010.07.012
                9628942
                20643162
                d3edb767-0841-44c9-8d14-570b23a1d027

                Elsevier has created a Monkeypox Information Center (https://www.elsevier.com/connect/monkeypox-information-center) in response to the declared public health emergency of international concern, with free information in English on the monkeypox virus. The Monkeypox Information Center is hosted on Elsevier Connect, the company's public news and information website. Elsevier hereby grants permission to make all its monkeypox related research that is available on the Monkeypox Information Center - including this research content - immediately available in publicly funded repositories, with rights for unrestricted research re-use and analyses in any form or by any means with acknowledgement of the original source. These permissions are granted for free by Elsevier for as long as the Monkeypox Information Center remains active.

                History
                : 28 April 2010
                : 30 June 2010
                : 12 July 2010
                Categories
                Short Communication

                Microbiology & Virology
                tnf, tumor necrosis factor,ct, threshold cycle,monkeypox virus west african strain,monkeypox virus congo basin strain,real-time pcr

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