| 中文摘要 | 第5-8页 |
| ABSTRACT | 第8-11页 |
| ABBREVIATIONS | 第16-19页 |
| Chapter I Metagenomic Analysis of Mosquito Viromes Isolated from Yunnan Provincein China reveals the Genes of Dengue and Zika Viruses | 第19-61页 |
| 1. NTRODUCTION | 第20-23页 |
| 2. MATERIALS AND METHODS | 第23-34页 |
| 2.1 Materials | 第23-24页 |
| 2.1.1 Experimental instruments | 第23页 |
| 2.1.2 Experimental reagents | 第23-24页 |
| 2.2 Methods | 第24-34页 |
| 2.2.1 Mosquito collection | 第24-25页 |
| 2.2.2 Grouping of Mosquito Samples | 第25-27页 |
| 2.2.3 Preparation of Mosquito Samples | 第27页 |
| 2.2.4 Extraction of Viral Nucleic Acid and Reverse Transcription | 第27-30页 |
| 2.2.5 RNA Degradation | 第30页 |
| 2.2.6 Purification of sscDNA | 第30页 |
| 2.2.7 Synthesis of Double-strand cDNA (dscDNA) | 第30-31页 |
| 2.2.8 Oligonucleotide Degradation | 第31页 |
| 2.2.9 Sequence-independent Single Primer Amplification (SISPA) | 第31页 |
| 2.2.10. Illumina Sequencing | 第31-32页 |
| 2.2.11 Computational Analysis | 第32页 |
| 2.2.12 Identification of Detected Virus | 第32-33页 |
| 2.2.13 Phylogenetic Analysis | 第33-34页 |
| 3. RESULTS | 第34-58页 |
| 3.1 Illumina Sequencing and the Virome of Mosquitoes | 第34-38页 |
| 3.2 Differential analysis of viral families in three samples | 第38-42页 |
| 3.3 PCR Identification of Metavirome Results | 第42-44页 |
| 3.4 PCR Amplification of Dengue Virus Gene | 第44-51页 |
| 3.5 PCR Amplification of Zika Virus Gene | 第51-55页 |
| 3.6 PCR Amplification of Japanese encephalitis virus Gene | 第55-58页 |
| 4. DISCUSSION | 第58-60页 |
| 5. CONCLUSIONS | 第60-61页 |
| Chapter Ⅱ Isolation, identification and Analysis of Japanese Encephalitis Virus fromMosquitoes in China, 2016 | 第61-98页 |
| 1. INTRODUCTION | 第62-65页 |
| 2. MATERIALS AND METHODS | 第65-77页 |
| 2.1 Design of universal primers | 第65页 |
| 2.2 Mosquitoes collection | 第65-66页 |
| 2.3 Extraction of viral RNA | 第66-67页 |
| 2.4 Detection of viruses | 第67页 |
| 2.5 Phylogenetic analysis for the full E sequences of JEV | 第67-69页 |
| 2.6 Cell culture | 第69页 |
| 2.7 Isolation of viruses | 第69页 |
| 2.8 Identification by indirect immunofluorescence assay (IFA) | 第69-70页 |
| 2.9 Observation of negative-stain electron microscopy | 第70页 |
| 2.10 Whole genome sequencing of newly isolated JEV | 第70-73页 |
| 2.10.1 Cell infection with the newly isolated JEV | 第70-71页 |
| 2.10.2 Viral RNA extraction and overlapping viral genome amplification | 第71-73页 |
| 2.11 Detection of viral titer | 第73页 |
| 2.12 Viral LDso detection on suckling mice | 第73-74页 |
| 2.13 Pathological changes in cerebrum of BALB/c suckling mice after inoculation with JEV | 第74页 |
| 2.14 Detection of viral titer of different passages | 第74页 |
| 2.15 The variability of envelope (E) gene of different passage viruses | 第74-75页 |
| 2.16 The variability of envelope gene of different passage viruses after BALB/c mice infection | 第75-77页 |
| 2.16.1 BALB/c mice infection with different passage viruses | 第75-76页 |
| 2.16.2 The variability of envelope gene of different passage viruses in BALB/c mice | 第76-77页 |
| 3. RESULTS | 第77-94页 |
| 3.1 Viral sequences detection and virus isolation | 第77页 |
| 3.2 CPE on BHK-21 cells | 第77-78页 |
| 3.3 Identification by indirect immunofluorescence assay (IFA) | 第78-79页 |
| 3.4 Negative-stain electron microscopy of JEV particles | 第79-80页 |
| 3.5 Phylogenetic analysis for the full E sequences of JEV | 第80-84页 |
| 3.6 Variation analysis for the full E sequences of JEV | 第84-86页 |
| 3.7 The viral titer and LDso of JEV-China/YN2016-1 | 第86-88页 |
| 3.8 Pathological changes in cerebrum of BALB/c suckling mice caused by JEV | 第88-89页 |
| 3.9 The viral titer of JEV-China/YN2016-1 of different passage viruses | 第89页 |
| 3.10 The variability of JEV-China/YN2016-1 E gene of different passage viruses | 第89-93页 |
| 3.11 The variability of envelope gene after BALB/c mice infection with different passage viruses | 第93-94页 |
| 4. DISCUSSION | 第94-97页 |
| 5. CONCLUSION | 第97-98页 |
| REFERENCES | 第98-118页 |
| 攻读博士学位期间的科研业绩 | 第118-120页 |
| 致谢 | 第120-121页 |
