海南疟疾流行区虫媒传染病的分子流行病学研究

As a tropical island, Hainan was one of the main malaria endemic area in China. Although no local malaria infection case had been reported in the past five years, but there were a number of FUO(fever of unknown origin)/anemia cases that microscope examination were negative happened in endemic areas every year. Because of the rich forest and bush, plentiful rainfall, a lot of various wild animals and vectou arthropods, many other known and unknown arthropod-borne infectious diseases (ABIDs) except malaria could be found in these areas. Due to the no systematic research on ABIDs in past times, the background and the pathogen spectrum of ABIDs were unclear in Hainan. Misdiagnosis and underdiagnosis were common occurences for the reasons without effective diagonsis and prevention methods for ABIDs. So, ABIDs caused the great threaten for the health of population and for construction of International Tourism Island and defence. In recent years, the emerging and re-emerging ABIDs were epidemic in the global or local area and caused serious public health problems. Therefor, the systematic survey of pathogen spectrum for ABIDs in malaria endemic areas is very important for control and prevent these diseases. In this project, we plan to do a molecular epidemiological survey on pathogens of ABID to the samples of FUO/amemia cases, major wild animal hosts and arthropods from endemic areas during the next two years using PCR/RT-PCR，Luminex high-throughput multi-analyte suspension array and high-throughput genome sequencing technology. The results of the project will obtain the systematic pathogen spectrum，time-space distribution characteristics of ABIDs ，genetic evolution characteristics of pathogens and construction of pathogen resources. This work is very important for providing the molecular epidemiological evidence of malaria elimination, providing scientific data and technological surpports for control of ABIDs in endemic areas in Hainan. It is also great significance for guaranteeing the health of the population, promoting the construction of International Tourism Island and national defense .

海南地处热带，曾是我国主要疟疾流行区之一，已5年无本地疟疾病例，但原疟疾流行区常年有镜检阴性的不明原因发热/贫血病人。流行区植被丰富，野生动物及节肢动物种类繁多，除疟疾外还存在多种已知及未知的虫媒传染病（ABIDs)。由于无系统的研究，流行区ABIDs本底及病原谱不清，缺乏有效的诊疗技术及防治措施，误诊、漏诊极为普遍，危害严重。本项目拟连续2年对流行区典型环境的不明原因发热/贫血患者及高危人群、主要动物宿主及节肢动物媒介进行采样，应用PCR/RT-PCR、高通量液相芯片及高通量基因组测序等技术开展ABIDs病原的分子流行病学调查，结果将初步获得海南疟疾流行区ABIDs的病原谱、分布的时空特征及病原体的遗传进化特征，构建病原资源库，是重要的基础工作，可为海南是否消除疟疾提供依据，为流行区ABIDs防治及研究提供科学数据及技术支撑，对保障民众健康、促进国际旅游岛及国防建设具有重要意义。

海南曾是我国主要疟疾流行区之一，已多年无本地疟疾病例，除疟疾外还存在多种已知及未知的虫媒传染病（ABIDs)，常年有不明原因发热/贫血病人。由于无系统的研究，流行区ABIDs 本底及病原谱不清，缺乏有效的诊疗技术及防治措施，误诊、漏诊极为普遍。本研究在项目资助下，应用PCR/RT-PCR及高通量基因组测序等技术对海南不明原因发热/贫血患者、主要动物宿主及节肢动物媒介样本开展ABIDs病原的分子流行病学研究，取得以下初步结果：1.建立了多种虫媒传染病PCR检测方法；2.应用PCR方法检测的海南不明原因发热（108例）/贫血（173例）患者血样Pv.、Pf.、Po.、Pm.，尚未发现阳性结果；3.应用PCR及病毒宏基因组测序技术检测6000余只蚊虫，未发现登革、寨卡等高致病性病毒；4.应用病毒宏基因组测序技术初步获得海南部分地区鼠类携带病毒性病原的本地数据，初期已分析鉴定出温州病毒海口株（HMU-virus）、Polyomavirus-HMU、Gemycirularvirus（GemyCV-HMU）等具有潜在致病性的病毒；5. 发现和鉴定了海南首例输入性发热伴血小板减少综合症病毒感染病例；6. 开展了海南机会致病性人兽共患肠道原虫的分子流行病学研究，发现在食蟹猴体内发现毕氏微孢子虫的7个新基因型 (分别命名为：HNM-I-HNM-VII)以及3个隐孢子虫新基因亚型，首次在豪猪、黄牛及黑山羊体内发现毕氏微孢子虫，在豪猪体内发现隐孢子虫，国内首次发现隐孢子虫旅行者株及毕氏微孢子虫Pongo2基因型；7. 对部分不明原因发热病人应用胶体金抗体检测及PCR方法恙虫东方次体检测，胶体金阳性率36.36%（24/66），PCR阳性率18.18%（12/66）；扩增产物测序Blast同源性分析，分别属恙虫东方次体Karp(7/12)、Gilliam（4/12）、Kawasaki（2/12）及TA763（1/12）株；8.已发表论文6篇，待发表5篇，申报专利2项，培养研究生4人，青年教师数人。项目初步获得海南不明原因发热/贫血患者、主要动物宿主及节肢动物媒介样本携带病原本底数据，可为海南ABIDs防治及研究提供科学数据及技术支撑，对保障民众健康、促进国际旅游岛及国防建设具有重要意义。

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