Establishment of LAMP assay for the detection of Necator Americanus in fecal samples
WEI Fu-rong, ZHU Hui-hui, JIA Xiao-kai, WANG Jun-yun
National Institute of Parasitic Diseases, Chinese Center for Disease Control and Prevention; WHO Collaborating Centre for Tropical Diseases; National Center for International Research on Tropical Diseases, Ministry of Science and Technology; Key Laboratory of Parasite and Vector Biology, Ministry of Health, Shanghai 200025, China
Abstract:Four primers based on ITS gene were designed to establish a loop-mediated isothermal amplification (LAMP) assay for rapid and effective detection of Necator Americanus eggs in feces. LAMP assays were performed to evaluate the specificity through detecting Schistosoma japonicum, Clonorchis sinensis, Trichuris trichura, Ascaris lumbricoides, Enterobius vermicularis. Good specificity for the LAMP assay with only specific amplifications on N. americanus among various species were found. The sensitivity of LAMP was performed on gradient dilutions of DNA samples of N. americanus,to evaluate the minimum detectable level. As results the minimum detectable level for the LAMP assay was 1.2 fg/mL. In addition, Kato-kazo, LAMP and conventional PCR were performed on 55 human fecal samples to compare the sensitivity and specificity. Regarding the fecal samples, the sensitivity of LAMP was 95.24% and 100%, respectively, comparing with Kato-kazo and PCR. And specificity of LAMP was 97.06% and 100%, respectively. In conclusion highly specific, sensitive and effective LAMP method has been developed for the detection of N. americanus eggs.
危芙蓉, 朱慧慧, 贾孝凯, 汪俊云. 环介导等温扩增法检测人粪样中美洲钩虫的方法建立[J]. 中国人兽共患病学报, 2020, 36(7): 530-533.
WEI Fu-rong, ZHU Hui-hui, JIA Xiao-kai, WANG Jun-yun. Establishment of LAMP assay for the detection of Necator Americanus in fecal samples. Chinese Journal of Zoonoses, 2020, 36(7): 530-533.