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.
Microbiology Services Division, Health Protection Agency, Porton Down, Salisbury, SP4 0JG, United Kingdom. barry.atkinson@hpa.org.uk
Barry
B
Atkinson
John
J
Chamberlain
Christopher H
CH
Logue
Nicola
N
Cook
Christine
C
Bruce
Stuart D
SD
Dowall
Roger
R
Hewson
D000818:Animals; D001483:Base Sequence; D002522:Chlorocebus aethiops; D017931:DNA Primers / Q000235:genetics; D003225:Hemorrhagic Fever Virus, Crimean-Congo / Q000235:genetics / Q000302:isolation & purification*; D006479:Hemorrhagic Fever, Crimean / Q000821:virology*; D006801:Humans; D008969:Molecular Sequence Data; D012367:RNA, Viral / Q000235:genetics / Q000302:isolation & purification; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D016415:Sequence Alignment; D013045:Species Specificity; D014709:Vero Cells
D017931:DNA Primers; D012367:RNA, Viral