John Chamberlain

Dengue virus serotype 3, Karachi, Pakistan.

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Development of a real-time RT-PCR assay for the detection of Crimean-Congo hemorrhagic fever virus.

Crimean-Congo hemorrhagic fever (CCHF) is a virulent tick-borne disease with a case fatality rate ranging from 10-50% for tick-borne transmission, and up to 80% for nosocomial transmission. Human cases have been reported in over 30 countries across Europe, Asia, and Africa. It appears to be spreading to new areas with several countries reporting their first human cases of CCHF disease within the past 10 years. We report a novel real-time RT-PCR assay designed to amplify a conserved region of the CCHF virus S segment. It is capable of detecting strains from all 7 groups of CCHF, including the AP92 strain that until recently represented a lineage of strains that were not associated with human disease. The limit of detection of the assay is 5 copies of target RNA, and the assay shows no cross-reactivity with other viruses from within the same genus, or with viruses causing similar human disease.

Development of an indirect ELISA method for the parallel measurement of IgG and IgM antibodies against Crimean-Congo haemorrhagic fever (CCHF) virus using recombinant nucleoprotein as antigen

Recombinant nucleoprotein from Crimean-Congo Haemorrhagic Fever (CCHF) virus was successfully derived from a baculovirus expression system and purified for use in a novel enzyme-linked immunosorbent assay (ELISA) diagnostic test. Comparable tests were used for detection of IgG and IgM antibodies, thus allowing efficient detection of both antibodies in parallel. The major benefits of the assay also included removing any requirement for polyclonal sera, thus eliminating variation in preparations and allowing standardisation between laboratories. The assay was successfully tested using a panel of positive sera supplied from samples identified as being positive in Turkey, Tajikistan and Kosovo and shown to be sensitive and specific. It is envisaged that this simple diagnostic ELISA for CCHF virus infection which removes the reliance on polyclonal antibody preparations, will be accessible to a wider range of laboratories enabling them to carry out routine diagnosis. This will improve the efficiency of diagnosis and subsequent management of infected patients.

Hazara virus infection is lethal for adult type I interferon receptor-knockout mice and may act as a surrogate for infection with the human-pathogenic Crimean–Congo hemorrhagic fever virus

Hazara virus (HAZV) is closely related to the Crimean-Congo hemorrhagic fever virus (CCHFV). HAZV has not been reported to cause human disease; work with infectious material can be carried out at containment level (CL)-2. By contrast, CCHFV causes a haemorrhagic fever in humans and requires CL-4 facilities. A disease model of HAZV infection in mice deficient in the type I interferon receptor is reported in this study. Dose-response effects were seen with higher doses, resulting in a shorter time to death and earlier detection of viral loads in organs. The lowest dose of 10 p.f.u. was still lethal in over 50 % of the mice. Histopathological findings were identified in the liver, spleen and lymph nodes, with changes similar to a recent mouse model of CCHFV infection. The findings demonstrate that inoculation of mice with HAZV may act as a useful surrogate model for the testing of antiviral agents against CCHFV.

Identification and analysis of Crimean-Congo hemorrhagic fever virus from human sera in Tajikistan

BACKGROUND Crimean-Congo hemorrhagic fever (CCHF) is a virulent tick-borne disease reported in more than 30 countries across Europe, Africa, and Asia. The disease is considered endemic in several Central Asian countries, including Tajikistan; however reports of human cases from these regions rarely reach the West. METHODS We analyzed all historical confirmed cases of CCHF in Tajikistan, mapping these reports against geographic data to assess risk areas. In addition, comprehensive analysis was undertaken on the 2010 human CCHF cohort to demonstrate effective methodologies for diagnosing this disease in-country. RESULTS These data show that CCHF is endemic in Tajikistan, and several large clusters have been recorded. Endemic foci of disease are localized to the southern region, with geographical factors such as altitude, monthly mean temperature, and monthly mean precipitation levels limiting establishment of tick vectors in other areas. Genomic analysis of viral RNA from a 2010 human case revealed high nucleotide homology (99%) to a strain isolated in Tajikistan in 1990. CONCLUSIONS CCHF is an important vector-borne and nosocomial pathogen in Tajikistan. The ability to rapidly detect cases using real-time RT-PCR shortly after admission in the hospital setting allows prompt implementation of barrier nursing techniques, therefore reducing onward transmission of the virus.

Genome Sequence of Ex-Afghanistan Crimean-Congo Hemorrhagic Fever Virus SCT Strain, from an Imported United Kingdom Case in October 2012

ABSTRACT Crimean-Congo hemorrhagic fever (CCHF) virus is a serious human pathogen causing severe hemorrhagic disease with a fatality rate of up to approximately 30%. We have determined the viral genomic sequence from an isolate that caused a fatal case of imported CCHF in the United Kingdom in October 2012.