〔目的〕通过对珠海注册养猪场及珠海口岸地区的蚊虫体内黄病毒的带毒率、宿主动物(猪群)及其人群进行血清黄病毒类(日本脑炎、登革热等)抗体水平的调查研究,为控制黄病毒在人群中的流行提供科学依据。〔方法〕采集养猪场猪血清、养猪场和口岸蚊标本、养猪场从业人员、入境的东南亚船员和城区部分发热病人血清,采用酶联免疫吸附试验(ELISA)法分别检测猪群和人群血清中的日本脑炎、登革热抗体;使用紫外灯诱蚊法在养猪场和口岸捕捉蚊类,经鉴定种类后,按20只蚊一组制成蚊悬液后,采用细胞培养、逆转录-聚合酶链反应(RT-PCR)以及荧光定量PCR技术检测蚊体内日本脑炎病毒、登革和西尼罗病毒等黄病毒的携带情况。〔结果〕1.猪血清日本脑炎病毒IgG抗体:结果已发表于《中国国境卫生检疫杂志》2007年第3期《珠海市部分注册猪场日本脑炎调查研究》。2.蚊媒带毒情况检测:2005年1月～2006年12月共捕获的2763只成蚊,经鉴定隶属4属8种,其中白纹伊蚊所占比例最高(32.75%),其次为致倦库蚊(26.06%)、三带喙库蚊(25.30%)、海滨库蚊(7.75%)、中华按蚊(3.84%)、骚扰阿蚊(3.18%),其他(常型曼蚊和巨型阿蚊等)(1.12%);在132组蚊研磨液的病毒细胞培养中,有7份标本出现CPE病变,使用黄病毒通用引物,RT-PCR反应能扩增出相应的阳性条带,其中白纹伊蚊组阳性3份,致倦库蚊组阳性3份,三带喙库蚊组阳性1份,但用日本脑炎病毒、登革病毒Ⅰ～Ⅳ型和西尼罗病毒等特异性引物,未能扩增出相应的阳性条带,说明可能存在着其它黄病毒类的病毒感染;采用荧光定量PCR法,对蚊悬液和细胞培养液日本脑炎、登革和西尼罗病毒进行检测,从1组海滨库蚊标本中检出日本脑炎病毒核酸阳性,但强度较弱,这与珠海地区是乙脑低发地区相一致。3.人群登革热和日本脑炎血清抗体检测:从东南亚的入境船员、本地发热病人和养猪场从业人员等血清标本511份中,登革病毒抗体IgG总阳性率为7.24%,其中以东南亚的入境船员的阳性率最高,为12.76%,没有检出登革病毒抗体IgM和日本脑炎IgM抗体。〔结论〕珠海地区存在日本脑炎的主要传播媒介,并且在蚊媒中携带有日本脑炎病毒;蚊标本细胞培养出现细胞病变和RT-PCR扩增出黄病毒基因的特异性片段,提示在捕获蚊标本中可能存在着其他病毒感染的可能;虽然养猪场猪群日本脑炎感染率不高,但存在着日本脑炎病毒的隐性感染或曾经感染过,提示不能够放松对乙型脑炎的预防控制工作,应采取积极的预防措施,防止人群和猪场日本脑炎的发生与流行。此外,研究结果表明东南亚船员有较高的登革热感染率,因此,在登革热流行期间,我国口岸应加强对来自东南亚国家交通工具员工和旅客的登革热的监测,以及有可能藏带、孳生蚊虫的废旧物品、集装箱等的检验检疫工作,以防止登革热的传入与流行。 [Objective] The aim of the study was to investigate the antibody levels of the serum flaviviruses (Japanese encephalitis and Dengue fever) in host animals (herds) and their populations by taking the virulent rate of flaviviruses in mosquitoes in Zhuhai registered pig farms and Zhuhai ports Research, to provide a scientific basis for controlling the prevalence of flaviviruses in the population. [Methods] The swine sera of swine farms, the mosquito samples of pig farms and ports, the employees of pig farms, the seamen arriving in Southeast Asia and some febrile patients in urban areas were collected. The swine herds were detected by enzyme - linked immunosorbent assay (ELISA) Japanese encephalitis and dengue antibody in the serum of the population; capture the mosquitoes in the pig farms and ports by using the UV lamp method; after being identified, the mosquito suspension is made into a group of 20 mosquitoes, and the cells are cultured and reversed PCR-RT-PCR and fluorescent quantitative PCR were used to detect the carriage of flavivirus such as Japanese encephalitis virus, dengue and West Nile virus in mosquitoes. [Results] 1. Porcine serum Japanese encephalitis virus IgG antibody: The results were published in “China Frontier Health and Quarantine Magazine” 2007 the third “Japanese registered encephalitis survey Zhuhai part of the farm.” 2. Mosquito-borne virus detection: A total of 2,763 adult mosquitoes were collected from January 2005 to December 2006, of which 8 species belong to 4 genera, of which Aedes albopictus accounted for the highest proportion (32.75%), followed by Culex pipiens quinquefasciatus (26.06%), Culex tritaeniorhynchus (25.30%), Cucurbita moschata (7.75%), Anopheles sinensis (3.84%), Alzheimer’s mosquito (3.18%), Giant mosquitoes, etc.) (1.12%). In the virus culture of 132 mosquito larvae, CPE lesions were found in 7 of the samples, and the corresponding positive bands were amplified by RT-PCR using flavivirus universal primers. Among them, 3 were positive in Aedes albopictus group, 3 were positive in Culex pipiens quinquefasciatus group and 1 in Culex tritaeniorhynchus group, but were positive for Japanese encephalitis virus, Dengue virus type Ⅰ ~ Ⅳ and West Nile virus The positive primers could not amplify the corresponding positive bands, indicating that there may be other viral infections of flaviviruses. Fluorescent quantitative PCR was used to detect the encephalitis, dengue and West Nile in mosquito suspension and cell culture medium The virus was detected from a group of Culex pipiens samples were detected positive for Japanese encephalitis virus nucleic acid, but the intensity is weak, which is consistent with areas of low prevalence of JE in Zhuhai. 3. Dengue dengue fever and Japanese encephalitis serum antibody detection: From 511 sera from Southeast Asia, sera from local febrile patients and employees in pig farms, the total positive rate of dengue virus IgG was 7.24%, of which, Southeast Asian The highest positive rate of entry crew was 12.76%, no dengue virus IgM and Japanese encephalitis IgM antibody were detected. [Conclusion] The major vectors of Japanese encephalitis exist in Zhuhai area and carry the Japanese encephalitis virus in the mosquito vectors. The cytopathic effect of cytopathic cells and the specific fragments of flavivirus genes amplified by RT-PCR are indicated in Zhuhai area. There may be other virus infections in the captured mosquito specimens. Although the infection rate of Japanese encephalitis in swine pig herds is not high, there is a latent infection or infection of Japanese encephalitis virus, suggesting that it is impossible to relax on type B Prevention and control of encephalitis should take proactive preventive measures to prevent the occurrence and prevalence of Japanese encephalitis in populations and farms. In addition, the findings show that Southeast Asian seafarers have a higher rate of dengue fever infection. Therefore, during the epidemic of dengue, monitoring of dengue fever among transport staff and travelers from Southeast Asian countries should be stepped up at the port of China and waste of piggybacks, breeding mosquitoes Goods, containers and other inspection and quarantine work in order to prevent the spread of imported and popular dengue.