A Rapid Method for Detection of Crimean Congo Hemorrhagic Fever Virus with Loop-Mediated Isothermal Amplification Assay

Abstract

Background: Crimean Congo hemorrhagic fever virus (CCHFV) is an extremely virulent pathogen, and ticks are a reservoir of the virus. One of the CCHFV endemic countries is Iran, where human cases of the infection have been reported in nearly all parts of the country. Objectives: In the present study, a reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay was designed for rapid detection of the small (S) RNA segment of CCHFV. Methods: A set of RT-LAMP primers from the highly conserved region of the S segment was designed for identification of CCHFV strains. The RT-LAMP, under isothermal conditions at 63°C, was performed without any superior equipment. Results: The sensitivity results indicated that RT-LAMP detected 62 fg of CCHFV RNA. A 100% agreement between the results of the RT-LAMP and RT-polymerase chain reaction (PCR) was observed. Conclusions: This method is more economical and practical than other molecular detection methods such as real-time PCR and nested PCR in developing countries. This method is suitable for rapid detection of this disease in rural hospitals and remote areas.