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Rapid Identification of Chikungunya and Dengue Virus by a Real-Time Reverse Transcription-Loop-Mediated Isothermal Amplification Method

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  • Department of Respiratory, Tangdu Hospital, Fourth Military Medical University, Xi'an, China; Guangdong Entry-Exit Inspection and Quarantine Bureau, Guangzhou, China; The State Key Laboratory of Respiratory Diseases, Guangzhou Medical University, Guangzhou, China; Department of Emergency, Tangdu Hospital, Fourth Military Medical University, Xi'an, China; South China University of Technology, Guangzhou, China

Both Chikungunya and Dengue virus belong to the acute arthropod-borne viruses. Because of the lack of specific symptoms, it is difficult to distinguish the two infections based on clinical manifestations. To identify and quantitatively detect Chikungunya and Dengue viruses, a real-time accelerated reverse-transcription-loop-mediated isothermal amplification (RT-LAMP) platform was developed, and 26-confirmed RNA samples, 42 suspects, and 18 healthy serum samples were evaluated by the method. The RT-polymerase chain reaction (PCR) and cDNA sequencing were used as references. The results showed that it could identify the Chikungunya and Dengue virus RNA correctly in all antibody-positive samples within 1 hour, without any cross-reactions. The virus load of the positive samples was quantitatively detected with a turbidimeter. The sensitivity was 100% and specificity was 95.25%. The findings indicate that the RT-LAMP is an effective method for rapid quantity detection of Chikungunya virus and Dengue virus in serum samples with convenient operation, high specificity, and high sensitivity.

Author Notes

* Address correspondence to Ziyao Mo, The State Key Laboratory of Respiratory Diseases, Guangzhou Medical University, Guangzhou, China. E-mail: ziyaomo@gmail.com

Financial support: This work was supported by State Key Laboratory of Respiratory Diseases (2007DA780154F0904), The Research Program General Administration of Quality Supervision, Inspection and Quarantine of the People's Republic of China (2010IK222), The Research Project of Guangdong Entry-Exit Inspection and Quarantine Bureau (2009GDK11).

Authors' addresses: Xi Lu and Faguang Jin, Department of Respiratory, Tangdu Hospital, Fourth Military Medical University, Xi'an, China, E-mails: xi.lu@mail.scut.edu.cn and jinfag@fmmu.edu.cn. Xiaobo Li, Guangdong Entry-Exit Inspection and Quarantine Bureau, Guangzhou, China, E-mail: lxb_1980@yahoo.cn. Ziyao Mo and Hongbo Zhao, The State Key Laboratory of Respiratory Diseases, Guangzhou Medical University, Guangzhou, China, E-mails: ziyaomo@gmail.com and zhaohongbolt@163.com. Boliang Wang, Department of Emergency, Tangdu Hospital, Fourth Military Medical University, Xi'an China, E-mail: wang.fmmu@gmail.com. Xiaoxiao Shan, School of Bioscience & Bioengineering, South China University of Technology, Guangzhou, China. Lei Shi, College of Light Industry and Food Sciences, South China University of Technology, Guangzhou, China, E-mail: leishi@scut.edu.cn.

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