Innovation
Focusing on the research and development and application of isothermal nucleic acid amplification technology
Recombinase-mediated isothermal amplification fluorescence assay for rapid detection of infective Oncomelania snails harboring Schistosoma japonicum—Jiangsu Institute of Parasitic Diseases
-
File size: 1.4MB
[Abstract] Objective: To establish a recombinase-mediated isothermal amplification fluorescence (RAA) assay for the rapid detection of Schistosoma japonicum-infected Oncomelania snails and to explore the optimal sample preparation method for Oncomelania snails. Methods: Oncomelania snail samples were divided into three groups, each containing seven subgroups. In six of these subgroups, 1, 2, 3, 4, or 5 infected snails were mixed into pools of 50 uninfected snails; in one subgroup, one infected snail was mixed into a pool of 100 uninfected snails. The three groups of snails were processed using three different methods: the crushed-shell nucleic acid extraction kit method, the crushed-shell crude nucleic acid extraction method, and the direct-crushed-shell crude nucleic acid extraction method. Both fluorescence RAA and PCR assays were performed on the extracted samples, and the results were compared. Results: A fluorescence RAA assay was established with an optimal reaction temperature of 39°C, enabling the detection of S. japonicum-infected Oncomelania snails within 30 minutes. After processing the pooled snail samples using the crushed-shell nucleic acid extraction kit method, the fluorescence RAA assay could detect as few as one infected snail mixed into a pool of 100 uninfected snails, whereas the PCR assay could detect at least one infected snail mixed into a pool of 50 uninfected snails. Using the crushed-shell crude nucleic acid extraction method, the fluorescence RAA assay could detect as few as one infected snail mixed into a pool of 100 uninfected snails, while the PCR assay could detect at least three infected snails mixed into a pool of 50 uninfected snails. After processing with the direct-crushed-shell crude nucleic acid extraction method, the fluorescence RAA assay could detect as few as 10 infected snails mixed into a pool of 50 uninfected snails, whereas the PCR assay could detect at least 10 infected snails mixed into a pool of 50 uninfected snails. Conclusion: We have successfully established a fluorescence RAA assay for the rapid detection of S. japonicum-infected Oncomelania snails in pooled snail samples. This assay features rapidity, high sensitivity, and ease of operation. The crushed-shell crude nucleic acid extraction method proved to be the optimal sample preparation approach for Oncomelania snails.
Product Inquiry
Contact: qt@qt-bio.com
Product Consultation:+86-510-85385531
Technical support:+86-18921157475
Address: No. 97,Xingye Building B,Linghu Avenue,Xiwu District, Wuxi City
WeChat Official Account
Official Official Account
www.300.cn