A one-step multiplex real-time RT-PCR for the universal detection of all currently known CCHFV genotypes
| Autor: | Martin H. Groschup, Ali Mirazimi, Miriam A. Sas, Marc Mertens, Martin Eiden, Serafeim C. Chaintoutis, Petra Emmerich, Ariel Vina-Rodriguez |
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| Rok vydání: | 2018 |
| Předmět: |
0301 basic medicine
Genotype 030231 tropical medicine Population Biology Real-Time Polymerase Chain Reaction Sensitivity and Specificity Arbovirus Virus 03 medical and health sciences 0302 clinical medicine Virology medicine Animals Seroprevalence Multiplex Seroconversion education education.field_of_study Reproducibility of Results medicine.disease biology.organism_classification 030104 developmental biology Hemorrhagic Fever Virus Crimean-Congo RNA Viral Hemorrhagic Fever Crimean Hyalomma Multiplex Polymerase Chain Reaction Crimean Congo hemorrhagic fever virus |
| Zdroj: | Journal of Virological Methods. 255:38-43 |
| ISSN: | 0166-0934 |
| Popis: | Crimean-Congo hemorrhagic fever (CCHF) is a fatal disease in humans, which is endemic in many countries of Africa, Southern Asia and Southeastern Europe. It is caused by the Crimean-Congo hemorrhagic fever virus (CCHFV), which is an arthropod-borne virus (arbovirus) transmitted by ixodid ticks, mainly of the genus Hyalomma. Animals like hares, hedgehogs, cattle, camels and small ruminants can become infected without developing clinical signs. Seroconversion occurs after a short viremia of up to two weeks, and thus seroprevalence studies in ruminants can be used to reveal risk areas for the human population. Virus detection by real-time quantitative reverse transcription polymerase chain reaction (RT-qPCR) is essential to prove an actual circulation of CCHFV in a country and is also used as diagnostic method for acute human CCHFV infections. In this study, a new universal one-step multiplex real-time RT-qPCR for the sensitive and specific detection of CCHFV is presented. For this purpose, 14 new primers and 2 probes were simultaneously used to detect RNAs representing all six CCHFV genotypes. Additionally, a GC-mirrored sequence within the synthetic RNAs enables the discrimination between true positive samples and unintentional laboratory contaminations. CCHFV negative samples from different animal species and ten different members of the order Bunyavirales were eventually tested to reveal the specificity of the new RT-qPCR. |
| Databáze: | OpenAIRE |
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