Innovative Technology

Innovation

Focusing on the research and development and application of isothermal nucleic acid amplification technology

重组酶介导的蓝氏贾第鞭毛虫特异性等温核酸扩增方法的建立及评价-江苏血防所

  • File size: 1.3MB

[Abstract] Objective: To establish a nucleic acid detection method for Giardia lamblia based on recombinase-aided amplification (RAA) technology and evaluate its detection sensitivity and specificity. Methods: The β-giardin gene of Giardia lamblia was selected as the target gene, and specific primers and a fluorescent probe were designed and synthesized to develop a fluorescence-based RAA assay system. The assay was validated by performing fluorescence RAA amplification using templates with varying copy numbers of a recombinant plasmid containing the B-giardin gene sequence, as well as genomic DNA from Giardia lamblia at different concentrations. Additionally, the specificity of the assay was assessed by amplifying genomic DNA from Giardia lamblia, Schistosoma japonicum, Clonorchis sinensis, Cryptosporidium parvum, Ascaris lumbricoides, Salmonella, and Shigella. Results: A reliable fluorescence RAA assay for Giardia lamblia was successfully established, enabling rapid and specific amplification of the target gene fragment under isothermal conditions (39°C) within 20 minutes. When using either the recombinant plasmid or Giardia lamblia genomic DNA as templates, the assay demonstrated a detection sensitivity of up to 10 copies/µL and 1 pg/µL, respectively. Furthermore, no amplification was observed when genomic DNA from Schistosoma japonicum, Clonorchis sinensis, Cryptosporidium parvum, Ascaris lumbricoides, Salmonella, or Shigella was used as the template, indicating excellent specificity. Conclusion: A simple, sensitive, and specific fluorescence RAA method has been developed for the detection of Giardia lamblia, offering a practical tool for molecular diagnostics in clinical and research settings.

Previous page

重组酶介导的华支睾吸虫特异性核酸等温扩增方法的建立及初步评价-江苏血防所

重组酶介导的日本血吸虫特异性基因片段核酸等温扩增检测方法的建立-江苏血防所

Next page

Previous page

重组酶介导的华支睾吸虫特异性核酸等温扩增方法的建立及初步评价-江苏血防所

Next page

重组酶介导的日本血吸虫特异性基因片段核酸等温扩增检测方法的建立-江苏血防所