声明
Chapter 1
1.1 Introduction
1.2 Distribution of Schistosomes
1.3 Life Cycle of Schistosoma
1.4 Pathogenesis of Schistosomiasis
1.5 Cell-Free DNA and Parasitic Infections
1.6 Source of cfDNA and Its Application in Diagnosis
1.7 Detection of Schistosoma in the Circulatory System
1.8 Problem and solution
Chapter 2 Recent advances in nucleic acid-based methods for detection of helminth infections and the perspective of biosensors for future development
2.1 Summary
2.2 Introduction
2.3 Nucleic acid-based methods for detection of helminth infections
2.3.1 PCR
2.3.2 Multiplex PCR
2.3.3 LAMP
2.3.4 Recombinase polymerase amplification (RPA)
2.4 Biosensors for nucleic acid biomarker detection
2.4.1 Electrochemical detection techniques
2.4.2 Optical detection techniques
2.5 Advantages and limitations of nucleic acid-based methods for detection of helminth infections
2.6 Conclusions
Chapter 3 Circulating cell free mitochondrial DNA fragments are useful markers for the early detection of Schistosoma japonicum infection
3.1 Abstract
3.2 Introduction
3.3 Material and Methods
3.3.1 Establishment of S. japonicum infection
3.3.2 Praziquantel(PZQ)treatment of infected mice and blood samples collection
3.3.3 Isolation of eggs from liver homogenate of infected mice and DNA extraction from eggs and adult S. japonicum
3.3.4 cfDNA extraction from infected host serum
3.3.5 Primer design and PCR amplification
3.3.6 Gene cloning in pMD 19 T vector and quantification of standard curve by qRT-PCR
3.3.7 Statistical analyses
3.4 Results
3.4.1 CfDNA extraction kits comparison
3.4.2 Reference Gene of S. japonicum detected in infected rabbit serum
3.4.3 Detection of S. japonicum cfDNA by PCR assay
3.4.4 Detection of S. japonicum cfDNA at different week of post infection
3.4.5 Detection of S. japonicum cfDNA in different number of cercariae of infection
3.4.6 Efficacy evaluation after chemotherapy
3.4.7 Characterization and copy number of target fragments
3.5 Discussion
3.6 Conclusion
Chapter 4 Detection of circulating cell-free nuclear DNA to diagnose Schistosoma japonicum infection
4.1 Abstract
4.2 Introduction
4.3 Material and Methods
4.3.1 Ethics statement
4.3.2 Establishment of S. japonicum infection, praziquantel treatment and collection of samples
4.3.4 Extraction of eggs from liver homogenate of infected mice and DNA extraction from eggs and adult S. japonicum infected host serum
4.3.5 PCR amplification
4.3.6 Gene cloning and standard curve construction by qRT-PCR for copy number detection
4.4 Results
4.5 Discussion
4.6 Conclusions
参考文献
致谢
CURRICULUM VITAE
中国农业科学院;
