Metagenomic sequencing for identifying pathogen-specific circulating DNAs and development of diagnostic methods for schistosomiasis
Jingyi Liu,
Xiaoxu Wang,
Fei Sheng,
Bikash R. Giri,
Shun Li,
Tianqi Xia,
Xuxin Li,
Guofeng Cheng
Affiliations
Jingyi Liu
Shanghai Tenth People’s Hospital, Tongji University School of Medicine, #500 Zhen-nan Road, Shanghai 200331, People’s Republic of China; Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Shanghai 200241, People’s Republic of China
Xiaoxu Wang
School of Biotechnology Jiangsu University of Science and Technology, Zhen Jiang 212100, People’s Republic of China
Fei Sheng
Shanghai Tenth People’s Hospital, Tongji University School of Medicine, #500 Zhen-nan Road, Shanghai 200331, People’s Republic of China
Bikash R. Giri
Shanghai Tenth People’s Hospital, Tongji University School of Medicine, #500 Zhen-nan Road, Shanghai 200331, People’s Republic of China
Shun Li
Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Shanghai 200241, People’s Republic of China
Tianqi Xia
Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Shanghai 200241, People’s Republic of China
Xuxin Li
Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Shanghai 200241, People’s Republic of China
Guofeng Cheng
Shanghai Tenth People’s Hospital, Tongji University School of Medicine, #500 Zhen-nan Road, Shanghai 200331, People’s Republic of China; Corresponding author
Summary: Timely diagnosis of Schistosoma infection, particularly in the early stage is crucial for identifying infected hosts and then taking effective control strategies. Here, metagenomic next-generation sequencing was used to identify pathogen-specific circulating DNAs (cDNAs) in the sera/plasma of New Zealand rabbits infected with S. japonicum, and the identified cDNAs were validated by PCR and qPCR. Loop-mediated isothermal amplification (LAMP)-based CRISPR-Cas12a and recombinase polymerase amplification-based lateral flow strip (RPA-LF) methods combined with the newly identified cDNA were developed to evaluate the potentials for diagnosing murine and human schistosomiasis. The results indicated that twenty-two cDNAs were identified. The developed LAMP-based CRISPR/Cas12a and RPA-LF methods showed a good potential for diagnosing murine or human schistosomiasis as early as 5 days of post-infection with 5 cercariae infection. In a word, S. japonicum specific cDNAs in circulation of infected hosts could be effective biomarkers for detecting Schistosoma infection particularly for early stages.
