Development and performance of recombinase-aided amplification (RAA) assay for detecting Schistosoma haematobium DNA in urine samples
Song Zhao,
Qiaoqiao Zhang,
Xinyao Wang,
Wei Li,
Saleh Juma,
Robert Berquist,
Jianfeng Zhang,
Kun Yang
Affiliations
Song Zhao
Jiangsu Institute of Parasitic Diseases, Key Laboratory on Technology for Parasitic Disease Prevention and Control, Ministry of Health, Jiangsu Provincial Key Laboratory on the Molecular Biology of Parasites, Wuxi, Jiangsu, China; Jiangnan University, Wuxi, Jiangsu, China
Qiaoqiao Zhang
Jiangsu Institute of Parasitic Diseases, Key Laboratory on Technology for Parasitic Disease Prevention and Control, Ministry of Health, Jiangsu Provincial Key Laboratory on the Molecular Biology of Parasites, Wuxi, Jiangsu, China; Department of Clinical Laboratory, The 904th Hospital of Joint Logistic Support Force of PLA, Wuxi, Jiangsu, 214044, China
Xinyao Wang
Jiangsu Institute of Parasitic Diseases, Key Laboratory on Technology for Parasitic Disease Prevention and Control, Ministry of Health, Jiangsu Provincial Key Laboratory on the Molecular Biology of Parasites, Wuxi, Jiangsu, China
Wei Li
Jiangsu Institute of Parasitic Diseases, Key Laboratory on Technology for Parasitic Disease Prevention and Control, Ministry of Health, Jiangsu Provincial Key Laboratory on the Molecular Biology of Parasites, Wuxi, Jiangsu, China
Saleh Juma
Ministry of Health of Zanzibar, P.O. Box 236, Zanzibar, United Republic of Tanzania
Robert Berquist
Ingerod, Brastad, Sweden (formerly with the UNICEF/UNDP/World Bank/WHO Special Programme for Research and Training in Tropical Diseases, World Health Organization, Geneva, Switzerland
Jianfeng Zhang
Jiangsu Institute of Parasitic Diseases, Key Laboratory on Technology for Parasitic Disease Prevention and Control, Ministry of Health, Jiangsu Provincial Key Laboratory on the Molecular Biology of Parasites, Wuxi, Jiangsu, China; Corresponding author.
Kun Yang
Jiangsu Institute of Parasitic Diseases, Key Laboratory on Technology for Parasitic Disease Prevention and Control, Ministry of Health, Jiangsu Provincial Key Laboratory on the Molecular Biology of Parasites, Wuxi, Jiangsu, China; Center for Global Health, School of Public Health, Nanjing Medical University, Nanjing, 211166, China; Jiangnan University, Wuxi, Jiangsu, China; Corresponding author. Jiangsu Institute of Parasitic Diseases, Key Laboratory on Technology for Parasitic Disease Prevention and Control, Ministry of Health, Jiangsu Provincial Key Laboratory on the Molecular Biology of Parasites, Wuxi, Jiangsu, China.
Rapid diagnosis of urogenital schistosomiasis caused by Schistosoma haematobium requires an accurate and timely assay, especially for low-intensity S. haematobium infection cases and in non-endemic areas. The mitochondrial cytochrome c oxidase 1 (cox1) gene fragment of S. haematobium was selected as detection target as this short fragment, which can be rapidly sequenced and yet possess good diagnostic resolution. A pair of primers and a fluorescent probe were designed according to the principle of recombinase-aided amplification (RAA), which was subsequently optimized and applied as an S. haematobium-specific RAA assay. Its diagnostic performance was validated for sensitivity and specificity in comparison to microscopy-based egg counting after urine filtration. The RAA assay could detect as little as 10 copies/μL of S. haematobium recombinant plasmid, and no cross-reactions were observed with S. mansoni, S. japonicum, Ancylostoma duodenale, Clonorchis sinensis, Echinococcus granulosus, or Ascaris lumbricoides. This test can be conducted at 39 °C and the whole RAA reaction can be completed within 20 min. The validation of the RAA assay showed that it had 100 % consistency with urine-egg microscopy, as it does not require an elaborate reading tool, is simple to use, and should be useful for field diagnostics and point-of-care applications.
