郑州和开封病人胃肠道主要寄生虫感染情况调查及蔬菜表面隐孢子虫卵囊洗脱方法的建立

发布时间：2018-06-04 13:41

本文选题：隐孢子虫 + 蔬菜　；参考：《河南农业大学》2013年硕士论文

【摘要】：隐孢子虫(Crycolosporidium)是一种专一的、寄生于胞内的寄生原虫，寄生于人和动物的胃肠道。能够引起严重的腹泻。食源性隐孢子虫病有明显的季节性，春季、夏季为易发季节。其传播途径非常广泛，可以以水源、食物、空气为介质进行传播，还能通过直接接触等途径传播，但是通过空气传播的报道非常少，通过以水源为介质传播的报道非常多，其在灌溉用水中被发现，是公认的世界范围内的水源性传播的病原体。食源性暴发的报道也非常多，1976年第一次有记录的人类感染隐孢子虫病暴发。此后很多国家相继报道该病的暴发。目前全球6大洲至少80个国家，，300多个地区报道了隐孢子虫病。近十几年来已经报道了多起食源性隐孢子虫病的暴发。隐孢子虫对人类健康造成了威胁，并且造成了不必要的经济损失。检测蔬菜表面的隐孢子虫卵囊，阻止其传播可以减免其造成的危害。Cook、Palmer和Joan等先后建立了水果、蔬菜表面检测隐孢子虫的方法。这些方法是基于4个基本阶段：从蔬菜中提取出卵囊；浓集并分离卵囊；使分离出的卵囊可视化；卵囊能在显微镜下识别。为了了解郑州开封两地患者肠道寄生虫感染情况，2012年7月4日至2012年8月31日期间，在郑州河南省人民医院，开封解放军第155医院和淮河医院采集的共2698份样品。采用卢戈氏碘染色法和饱和蔗糖漂浮法对所有样品进行了检查。共发现了5种肠道寄生虫，分别是环孢子虫、贾第虫、阿米巴原虫、线虫和钩虫。寄生虫阳性粪便样品29例，肠道总感染率为1.07%（29/2698）。其中环孢子虫和贾第虫感染率并列最高，均为0.4%（11/2698），其次为阿米巴原虫肠道感染，感染率为0.19%(5/2698)，线虫和钩虫，感染率都为0.04%(1/2698)。此次调查没有发现混合感染，也没有发现隐孢子虫。为了建立一种能检测蔬菜表面隐孢子虫的方法，用不同数量梯度的隐孢子虫卵囊对蔬菜进行接种，分别设置为5×101个、5×102个、5×103个、5×104个；再对蔬菜进行洗脱，采用不同的洗涤剂，分别为Alconox洗洁精、雕牌洗洁精、立白洗洁精、白猫洗洁精和去离子水；对卵囊的收集，采用反复离心收集沉淀法；被接种的蔬菜的用量也被加以对比，最后基于隐孢子虫18S rRNA基因位点对洗脱后的卵囊进行PCR扩增，建立了一个系统的检测方法。结果表明用50g西葫芦、15g生菜时，卵囊最小洗脱量均为5×101个；洗脱时间10min和20min结果无明显差异；接种12h和接种24h结果无明显差异；不同洗涤剂（Alconox洗洁精、雕牌洗洁精、立白洗洁精、白猫洗洁精、去离子水）洗脱结果无明显差异。成功建立一种能够检测农产品表面污染隐孢子虫卵囊的检测技术，为进一步开展病原溯源研究提供了技术支持。
[Abstract]:Cryptosporidium (Crycolosporidium) is a specific parasitic protozoa, parasitic on the gastrointestinal tract of humans and animals. Can cause severe diarrhea. Food-borne cryptosporidiosis has obvious seasonality, spring and summer are susceptible to occurrence. It can be spread through water, food and air as a medium, and through direct contact. However, there are very few reports of air transmission, and a lot of reports spread through water source as the medium. It is found in irrigation water and is recognized as a worldwide water-borne pathogen. Food-borne outbreaks have also been reported, with the first recorded human outbreak of Cryptosporidiosis in 1976. Since then, many countries have reported outbreaks of the disease. Cryptosporidiosis has been reported in more than 300 regions in at least 80 countries on 6 continents. Outbreaks of food-borne Cryptosporidiosis have been reported in recent decades. Cryptosporidium poses a threat to human health and causes unnecessary economic losses. The method of detecting Cryptosporidium oocysts on vegetable surface and preventing them from spreading can reduce the damage caused by Cryptosporidium parvum. Cooku Palmer and Joan have established a method for detecting Cryptosporidium on vegetable surface. These methods are based on four basic stages: extraction of egg sac from vegetables; concentration and separation of egg sac; visualization of separated egg sac; egg sac can be recognized under microscope. From July 4, 2012 to August 31, 2012, 2698 samples were collected from the 155th Hospital of Kaifeng PLA and Huaihe Hospital in Henan Provincial people's Hospital of Zhengzhou. All samples were examined by Lugo's iodine staining and saturated sucrose floatation. Five kinds of intestinal parasites were found, namely cyclosporium, giardia, amoeba protozoa, nematode and hookworm. The total intestinal infection rate of 29 cases of parasite positive fecal samples was 1.077% 2698%. The infection rates of Cyclosporidium and Giardia were the highest (0.411 / 2698), followed by amoebic protozoa (0.19 / 2698), nematode and hookworm (0.04 / 2698). No mixed infection was found in the investigation, nor was Cryptosporidium found. In order to establish a method to detect Cryptosporidium spore on vegetable surface, the vegetable was inoculated with different quantity gradient Cryptosporidium oocysts, which were 5 脳 10 1, 5 脳 10 2, 5 脳 10 3, 5 脳 10 4, and then eluted with different detergents. They are Alconox detergent, Diaodai detergent, Libai detergent, White Cat detergent and deionized Water; the egg sac is collected by repeated centrifugation and precipitation; and the amount of vegetables inoculated is also compared. Finally, based on the 18s rRNA locus of Cryptosporidium, a systematic detection method was established by PCR amplification of the eluted oocysts. The results showed that the minimum amount of egg sac elution was 5 脳 101 when 50 g squash was used in 15 g lettuce, there was no significant difference between 10min and 20min in elution time, there was no significant difference between inoculation at 12 h and 24 h after inoculation. There was no significant difference in the elution results between Li Bai detergent, white cat detergent and deionized water. A technique for detecting Cryptosporidium oocysts in agricultural products was successfully established, which provided technical support for the further study of pathogen traceability.
【学位授予单位】：河南农业大学
【学位级别】：硕士
【学位授予年份】：2013
【分类号】：S436.3;R531.5

【相似文献】