運(yùn)用重組隱孢子蟲特異性抗原對(duì)中國(guó)社區(qū)人群隱孢子蟲感染的血清流行病學(xué)研究
發(fā)布時(shí)間：2018-07-21 09:47
【摘要】： 隱孢子蟲(Cryptosporidium)為一種主要寄生于宿主消化道上皮細(xì)胞，引起以腹瀉為主要臨床癥狀的人獸共患寄生原蟲。免疫功能正常者，腹瀉可自限，但免疫功能低下者，，尤其是嬰幼兒和艾滋病患者，一旦感染則病情嚴(yán)重，遷延不愈，甚至危及生命。 近年來(lái)，在美國(guó)和加拿大等國(guó)家因水源污染引起了大規(guī)模的隱孢子蟲病爆發(fā)流行，造成了嚴(yán)重的經(jīng)濟(jì)損失，發(fā)達(dá)國(guó)家政府已經(jīng)意識(shí)到監(jiān)測(cè)隱孢子蟲對(duì)水源的污染以及在環(huán)境中分布情況的重要性。特別是近年來(lái)艾滋病蔓延，各種環(huán)境和職業(yè)性有害因素導(dǎo)致機(jī)體免疫抑制的情況廣泛存在，使得隱孢子蟲感染和傳播成為可能。因此，隱孢子蟲病的研究受到廣泛關(guān)注。 以往，隱孢子蟲病的診斷和流行病學(xué)調(diào)查依賴于通過(guò)鏡檢發(fā)現(xiàn)糞便中的隱孢子蟲卵囊，其中改良抗酸染色法因其特異性較好而被廣泛使用。但是，作為“金標(biāo)準(zhǔn)”的糞便檢查不僅工作量大，現(xiàn)場(chǎng)應(yīng)用時(shí)人群的依從性差，而且缺乏足夠的敏感性。 免疫學(xué)診斷方法因其快捷、簡(jiǎn)便易行等優(yōu)點(diǎn)，愈來(lái)愈受到重視。特異性抗體檢測(cè)是一種間接診斷方法，并不能肯定地對(duì)個(gè)案作出有或無(wú)的回答。但相關(guān)研究表明，血清學(xué)群體特征可以在一定程度上反映相應(yīng)流行區(qū)的流行狀態(tài)，并為疾病的防控提供重要資料。 本研究選擇了兩個(gè)社區(qū)經(jīng)濟(jì)條件、日用水源及健康衛(wèi)生狀況存在顯著差異的中國(guó)農(nóng)村社區(qū)，收集其全人群的戶口信息，同時(shí)采集血清，運(yùn)用CP23重組蛋白為抗原的間接ELISA法，定量檢測(cè)人群隱孢子蟲CP23抗原特異的抗體水平，并根據(jù)陽(yáng)性判斷值，估計(jì)隱孢子蟲在上述兩個(gè)社區(qū)的流行狀態(tài)及其影響因素。 本研究獲得結(jié)果如下： 1．成功制備了純化的CP23蛋白，并在本室建立了以此蛋白為抗原的間接ELISA法用于檢測(cè)隱孢子蟲特異性抗體。 2．首次對(duì)中國(guó)農(nóng)村人群的隱孢子蟲感染狀態(tài)進(jìn)行血清流行病學(xué)調(diào)查。浙江麻車村人群隱孢子蟲感染的血清抗體陽(yáng)性率為32.16％(173／538)，江西駕湖村為70.20％(688／980)，兩社區(qū)血清抗體陽(yáng)性率差異顯著(P=0.000)。 3．對(duì)浙江及江西兩地隱孢子蟲感染人群血清抗體陽(yáng)性率與相關(guān)變量的廣義估計(jì)模型分析表明在社區(qū)內(nèi)部，教育程度、家庭收入和有無(wú)血吸蟲感染對(duì)隱孢子蟲抗體陽(yáng)性率沒(méi)有影響，而性別和年齡是影響隱孢子蟲抗體陽(yáng)性率的重要因素。人群隱孢子蟲感染血清抗體陽(yáng)性率存在性別差異。浙江麻車村人群，在6-10歲年齡組女性低于男性，21-30歲年齡組男性與女性的血清抗體陽(yáng)性率接近，而其余各年齡組女性隱孢子蟲感染血清抗體陽(yáng)性率高于男性。男性在各年齡組血清抗體陽(yáng)性率相當(dāng)，而女性的隱孢子蟲血清抗體陽(yáng)性率隨著年齡增長(zhǎng)而呈上升的趨勢(shì)，在41-50歲年齡組達(dá)到最高值48.05％。江西駕湖村人群各年齡組女性隱孢子蟲感染血清抗體陽(yáng)性率均高于男性。且男性的隱孢子蟲感染血清抗體陽(yáng)性率有隨著年齡增長(zhǎng)而上升的趨勢(shì)，在41-50歲年齡組達(dá)到最高值(73.86％)。而女性各年齡組血清陽(yáng)性率相當(dāng)，無(wú)此趨勢(shì)。 4．兩個(gè)地區(qū)相比，江西人群的家庭收入顯著低于浙江人群(P=0.000)，江西人群的經(jīng)濟(jì)收入多在7000-10000元／年，而95％的浙江人群家庭年收入在16000-20000元及以上。通常，經(jīng)濟(jì)水平的差異會(huì)造成衛(wèi)生條件的不同。隱孢子蟲病是一種經(jīng)口傳播的腸道寄生蟲病，它的傳播途徑與大部分腸道蠕蟲相似，該寄生蟲的感染與衛(wèi)生條件有關(guān)，這些結(jié)果提示了家庭收入可能與兩地隱孢子蟲血清陽(yáng)性率的差異相關(guān)。 5．利用網(wǎng)絡(luò)數(shù)據(jù)庫(kù)，包括最新的ABCpred服務(wù)器對(duì)微小隱孢子蟲SA35和SA40蛋白的B細(xì)胞抗原表位進(jìn)行預(yù)測(cè)。隱孢子蟲SA35蛋白的103～115和129～146以及SA40蛋白的77～89、127～136、156～174和200～209等區(qū)段為B細(xì)胞表位。所得表位為這兩種蛋白今后應(yīng)用于合成肽檢測(cè)血清中抗體、制備相應(yīng)的抗體、聯(lián)合CP23抗原發(fā)展高特異性和敏感性的診斷系統(tǒng)等提供了理論基礎(chǔ)。
[Abstract]:Cryptosporidium (Cryptosporidium) is a kind of parasitic protozoa which mainly parasitic on the epithelial cells of the digestive tract of the host, causing the parasitic protozoa with diarrhea as the main clinical symptom. The immune function is normal, the diarrhea can be self limiting, but the low immune function, especially in infants and AIDS patients, is serious, not more prolonged and even endangered once the infection is infected. Life.
In recent years, a large scale of Cryptosporidium disease caused by water pollution in the United States and Canada has caused a massive outbreak of Cryptosporidium disease, causing serious economic losses. The government of developed countries has realized the importance of monitoring the pollution of Cryptosporidium to the water source and the importance of its distribution in the environment. It is possible that the infection and transmission of Cryptosporidium is possible because of the widespread harmful factors that cause the immune suppression of the body. Therefore, the study of Cryptosporidium is widely concerned.
In the past, the diagnosis and epidemiological investigation of cryptosporidiosis depended on the discovery of Cryptosporidium oocyst in feces by microscopic examination. The improved anti acid staining method was widely used. However, the fecal examination, as the "gold standard", was not only a large amount of work, but poor adherence to the population in the field, and it was not enough. Sensitivity.
Immunological diagnosis has been paid more and more attention because of its advantages, such as quick, easy and easy to do. Specific antibody detection is an indirect diagnostic method, and it is not sure to make any answer to the case. But the related research shows that the serological characteristics can reflect the epidemic state of the corresponding epidemic area to a certain extent and for the disease. Prevention and control provide important information.
In this study, we selected two community economic conditions, Chinese rural communities with significant differences in daily use of water and health and health conditions, collecting their population information, collecting serum and using the indirect ELISA method of CP23 recombinant protein as antigen to detect the specific antibody level of CP23 antigen of Cryptosporidium, and the positive judgment. The prevalence and influencing factors of Cryptosporidium in these two communities were estimated.
The results of this study are as follows:
1. the purified CP23 protein was successfully prepared, and an indirect ELISA method was established in this laboratory to detect Cryptosporidium specific antibodies.
2. the first seroepidemiological survey of Cryptosporidium infection in Chinese rural population was carried out. The positive rate of serum antibody of Cryptosporidium infection in Zhejiang mache village was 32.16% (173 / 538), 70.20% (688 / 980) in Jiangxi driving Lake Village, and significant difference in the positive rate of serum antibody in the two community (P=0.000).
3. a generalized estimation model of the positive rate of serum antibody and related variables of Cryptosporidium infective people in Zhejiang and Jiangxi showed that the degree of education, family income and infection without schistosomiasis had no effect on the positive rate of Cryptosporidium antibody in the community, and sex and age were important factors affecting the positive rate of Cryptosporidium. The positive rate of serum antibody of Cryptosporidium infection in the population of the population of Zhejiang Ma Che village was lower than that of male in the age group of 6-10 years old. The positive rate of serum antibody was close to the male and female in 21-30 years old age group, but the positive rate of serum antibody in the other age groups was higher than that of the male. The positive rate was equal, but the positive rate of serum antibody of Cryptosporidium in women increased with age, and the positive rate of serum antibody positive rate of Cryptosporidium infecting in the age group of 41-50 years old was higher than that of men in the age group of 41-50 years old, and the positive rate of serum antibody of Cryptosporidium infection in male was higher than that of men. The rising trend of age was the highest in the 41-50 year old group (73.86%).
4. compared with the two regions, the family income of the Jiangxi population was significantly lower than that of the Zhejiang population (P=0.000), and the economic income of the Jiangxi population was 7000-10000 yuan per year, while the annual income of the 95% Zhejiang population was 16000-20000 yuan and above. Generally, the difference in the economic level would cause the difference of health conditions. The transmission pathway of intestinal parasitosis is similar to that of most intestinal worms. The infection of the parasite is related to the hygienic conditions. These results suggest that the family income may be related to the difference in the positive rate of the sera of Cryptosporidium.
5. using the network database, including the latest ABCpred servers, the B cell epitopes of the SA35 and SA40 proteins of Cryptosporidium parvum are predicted. The 103~115 and 129~146 of Cryptosporidium SA35 protein and 77~89127 to 136156~174 and 200~209 of the protein are B cell epitopes. The epitopes are the future of these two proteins It provides a theoretical basis for the detection of antibodies in serum by synthetic peptides, the preparation of corresponding antibodies, and the development of highly specific and sensitive diagnostic systems combined with CP23 antigen.
【學(xué)位授予單位】：南京醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2007
【分類號(hào)】：R392;R531.5
本文編號(hào)：2135130
[Abstract]:Cryptosporidium (Cryptosporidium) is a kind of parasitic protozoa which mainly parasitic on the epithelial cells of the digestive tract of the host, causing the parasitic protozoa with diarrhea as the main clinical symptom. The immune function is normal, the diarrhea can be self limiting, but the low immune function, especially in infants and AIDS patients, is serious, not more prolonged and even endangered once the infection is infected. Life.
In recent years, a large scale of Cryptosporidium disease caused by water pollution in the United States and Canada has caused a massive outbreak of Cryptosporidium disease, causing serious economic losses. The government of developed countries has realized the importance of monitoring the pollution of Cryptosporidium to the water source and the importance of its distribution in the environment. It is possible that the infection and transmission of Cryptosporidium is possible because of the widespread harmful factors that cause the immune suppression of the body. Therefore, the study of Cryptosporidium is widely concerned.
In the past, the diagnosis and epidemiological investigation of cryptosporidiosis depended on the discovery of Cryptosporidium oocyst in feces by microscopic examination. The improved anti acid staining method was widely used. However, the fecal examination, as the "gold standard", was not only a large amount of work, but poor adherence to the population in the field, and it was not enough. Sensitivity.
Immunological diagnosis has been paid more and more attention because of its advantages, such as quick, easy and easy to do. Specific antibody detection is an indirect diagnostic method, and it is not sure to make any answer to the case. But the related research shows that the serological characteristics can reflect the epidemic state of the corresponding epidemic area to a certain extent and for the disease. Prevention and control provide important information.
In this study, we selected two community economic conditions, Chinese rural communities with significant differences in daily use of water and health and health conditions, collecting their population information, collecting serum and using the indirect ELISA method of CP23 recombinant protein as antigen to detect the specific antibody level of CP23 antigen of Cryptosporidium, and the positive judgment. The prevalence and influencing factors of Cryptosporidium in these two communities were estimated.
The results of this study are as follows:
1. the purified CP23 protein was successfully prepared, and an indirect ELISA method was established in this laboratory to detect Cryptosporidium specific antibodies.
2. the first seroepidemiological survey of Cryptosporidium infection in Chinese rural population was carried out. The positive rate of serum antibody of Cryptosporidium infection in Zhejiang mache village was 32.16% (173 / 538), 70.20% (688 / 980) in Jiangxi driving Lake Village, and significant difference in the positive rate of serum antibody in the two community (P=0.000).
3. a generalized estimation model of the positive rate of serum antibody and related variables of Cryptosporidium infective people in Zhejiang and Jiangxi showed that the degree of education, family income and infection without schistosomiasis had no effect on the positive rate of Cryptosporidium antibody in the community, and sex and age were important factors affecting the positive rate of Cryptosporidium. The positive rate of serum antibody of Cryptosporidium infection in the population of the population of Zhejiang Ma Che village was lower than that of male in the age group of 6-10 years old. The positive rate of serum antibody was close to the male and female in 21-30 years old age group, but the positive rate of serum antibody in the other age groups was higher than that of the male. The positive rate was equal, but the positive rate of serum antibody of Cryptosporidium in women increased with age, and the positive rate of serum antibody positive rate of Cryptosporidium infecting in the age group of 41-50 years old was higher than that of men in the age group of 41-50 years old, and the positive rate of serum antibody of Cryptosporidium infection in male was higher than that of men. The rising trend of age was the highest in the 41-50 year old group (73.86%).
4. compared with the two regions, the family income of the Jiangxi population was significantly lower than that of the Zhejiang population (P=0.000), and the economic income of the Jiangxi population was 7000-10000 yuan per year, while the annual income of the 95% Zhejiang population was 16000-20000 yuan and above. Generally, the difference in the economic level would cause the difference of health conditions. The transmission pathway of intestinal parasitosis is similar to that of most intestinal worms. The infection of the parasite is related to the hygienic conditions. These results suggest that the family income may be related to the difference in the positive rate of the sera of Cryptosporidium.
5. using the network database, including the latest ABCpred servers, the B cell epitopes of the SA35 and SA40 proteins of Cryptosporidium parvum are predicted. The 103~115 and 129~146 of Cryptosporidium SA35 protein and 77~89127 to 136156~174 and 200~209 of the protein are B cell epitopes. The epitopes are the future of these two proteins It provides a theoretical basis for the detection of antibodies in serum by synthetic peptides, the preparation of corresponding antibodies, and the development of highly specific and sensitive diagnostic systems combined with CP23 antigen.
【學(xué)位授予單位】：南京醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2007
【分類號(hào)】：R392;R531.5
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