Development of a portable reverse transcription loop‐mediated isothermal amplification system to detect the E1 region of Chikungunya virus in a cost‐effective manner
Benjawan Saechue, Naganori Kamiyama, Yinan Wang, Chiaki Fukuda, Kei Watanabe, Yasuhiro Soga, Mizuki Goto, Astri Dewayani, Shimpei Ariki, Haruna Hirose, Sotaro Ozaka, Nozomi Sachi, Shinya Hidano, Khaledul Faisal, Rajashree Chowdhury, Md Anik Ashfaq Khan, Faria Hossain, Prakash Ghosh, Tahmina Shirin, Dinesh Mondal, Kazunari Murakami, Takashi Kobayashi
Abstract
Chikungunya fever is a mosquito-borne disease cause of persistent arthralgia. The current diagnosis of Chikungunya virus (CHIKV) relies on a conventional reverse transcription polymerase chain reaction assay. Reverse transcription loop-mediated isothermal amplification (RT-LAMP) is a rapid and simple tool used for DNA-based diagnosis of a variety of infectious diseases. In this study, we established an RT-LAMP system to recognize CHIKV by targeting the envelope protein 1 (E1) gene that could also detect CHIKV at a concentration of 8 PFU without incorrectly detecting other mosquito-borne viruses. The system also amplified the E1 genome in the serum of CHIKV-infected mice with high sensitivity and specificity. Moreover, we established a dry RT-LAMP system that can be transported without a cold chain, which detected the virus genome in CHIKV-infected patient samples with high accuracy. Thus, the dry RT-LAMP system has great potential to be applied as a novel CHIKV screening kit in endemic areas.