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Focusing on the research and development and application of isothermal nucleic acid amplification technology
Recombinase-mediated isothermal amplification of nucleic acids... Evaluation of the efficacy of Oncomelania snails infected with Schistosoma—Ye Yuying
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[Abstract] Objective: To evaluate the efficacy of a recombinase-mediated isothermal amplification (RAA) fluorescence assay for detecting infective snails of Schistosoma japonicum. Methods: A pooled-sample approach was used. Every 50 snails constituted one test sample; negative samples contained no infective snails, while positive samples contained varying numbers of infective snails. Ten negative samples and 10 positive samples each containing 12 and 3 infective snails, respectively, were randomly divided into groups and tested by blinded fluorescence RAA assay. The results of the fluorescence RAA assay were compared with those obtained by the miracidial hatching method, which served as the gold standard, to calculate the sensitivity, specificity, Youden’s index, and concordance rate of the fluorescence RAA assay. Additionally, five negative snail samples and five positive samples each containing 1, 2, and 3 infective snails were randomly grouped, and paired-design testing was conducted, in which the same samples were tested blindly by both crushing microscopy and fluorescence RAA assay, and the results were compared. Results: The fluorescence RAA assay detected 29 out of 30 positive samples as positive, yielding a sensitivity of 96.67%. Among the 10 negative samples, 8 were correctly identified as negative, resulting in a specificity of 80.00%. The Youden’s index was 0.77, and repeated testing of the same samples 10 times showed a 100% concordance rate. There was no statistically significant difference between the results of the fluorescence RAA assay and those of crushing microscopy for detecting infective snails (P = 0.00 > 0.05). The concordance rates between the two methods were 95.00% (19/20) and 90.00% (18/20), respectively. Conclusion: The fluorescence RAA assay demonstrates good detection performance for infective snails of Schistosoma japonicum and holds promising potential for application in the screening of such snails in areas endemic for Japanese schistosomiasis.
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