本文選題：豬偽狂犬病 + gE-ELISA��； 參考：《福建農(nóng)林大學(xué)》2015年碩士論文

【摘要】：目的：利用gE-ELISA和gB-ELISA試劑盒評(píng)估分析福建省南平市延平區(qū)兩個(gè)規(guī)模化豬場(chǎng)的豬偽狂犬野毒感染率,并建立PCR診斷方法對(duì)兩豬場(chǎng)豬偽狂犬野毒的垂直傳播風(fēng)險(xiǎn)進(jìn)行分析。方法：對(duì)2014年11月～12月份南平市延平區(qū)兩個(gè)規(guī)�；i場(chǎng)進(jìn)行豬偽狂犬gE抗體及gB抗體檢測(cè),并對(duì)結(jié)果進(jìn)行統(tǒng)計(jì)分析,評(píng)估兩個(gè)規(guī)�；i場(chǎng)的野毒感染陽(yáng)性率。將統(tǒng)計(jì)結(jié)果與2013年南平市延平區(qū)流行病疫情調(diào)查結(jié)果進(jìn)行對(duì)比分析,以評(píng)估2014年這兩個(gè)豬場(chǎng)豬偽狂犬病毒(PRV)的流行進(jìn)展�；趃E基因建立檢測(cè)豬偽狂犬(PR)野毒的PCR診斷方法,并分析建立的PCR方法的特異性、靈敏性、重復(fù)性以及與國(guó)家標(biāo)準(zhǔn)的相符性。使用本試驗(yàn)建立的PCR方法檢測(cè)兩規(guī)�；i場(chǎng)和延平區(qū)12個(gè)豬場(chǎng)的gE抗體陽(yáng)性豬群的臍帶血以及公豬精液,統(tǒng)計(jì)對(duì)比兩豬場(chǎng)臍帶血和精液的PCR陽(yáng)性檢出率與12個(gè)豬場(chǎng)的平均陽(yáng)性率,評(píng)估兩規(guī)�；i場(chǎng)的垂直傳播風(fēng)險(xiǎn)壓力。結(jié)果與分析：A豬場(chǎng)共抽樣353份血清,B豬場(chǎng)共抽樣267份血清,gE抗體檢測(cè)結(jié)果表明,A、B兩場(chǎng)所檢測(cè)的豬群中除公豬外,其他所有豬群均檢出gE抗體,總陽(yáng)性率分別為：51.08%、58.80%,說(shuō)明了兩場(chǎng)均是PR野毒的陽(yáng)性場(chǎng)。兩場(chǎng)公豬的gE抗體陽(yáng)性率為0.00%,屬于陰性豬群,說(shuō)明了公豬群未被PR野毒感染。A、B豬場(chǎng)的母豬gE抗體陽(yáng)性率分別為：53.87%、46.15%；30～40日齡保育豬的gE抗體陽(yáng)性率分別為：26.09%、51.16%,加上可疑數(shù)的比例后分別為：31.89%、51.16%；60日齡保育豬的gE抗體陽(yáng)性率分別為：45.45%、65.96%；而90～120日齡豬群gE抗體陽(yáng)性率高達(dá)90%以上,由此推斷在轉(zhuǎn)舍(保育舍轉(zhuǎn)至肥育舍)時(shí)該階段豬群被PR野毒感染。gB抗體檢測(cè)結(jié)果表明：兩場(chǎng)的gB總抗體陽(yáng)性率分別為96.87%、96.84%；兩個(gè)豬場(chǎng)公豬的gB抗體陽(yáng)性率都為100.00%；母豬的gB抗體陽(yáng)性率分別為93.4%、100.00%,抗體水平高且離散度低,有可能是疫苗免疫和PR野毒感染的綜合作用；30-40日齡保育豬的gB抗體陽(yáng)性率分別為94.20%、97.67%,且整齊度高,因此可推斷除了受母源抗體的影響,兩豬場(chǎng)的首免效果好,而抗體水平A豬場(chǎng)高于B豬場(chǎng)；兩豬場(chǎng)60日齡至120日齡仔豬血清的gB抗體S/N值持續(xù)降低,即抗體水平升高,而此階段母源抗體已幾乎消退,gE抗體也不降反升,因此推斷60日齡尤其是120日齡轉(zhuǎn)舍時(shí)豬群受野毒感染。2014年A場(chǎng)gE抗體陽(yáng)性率平均值為51.08%,B場(chǎng)gE抗體陽(yáng)性率平均值為58.80%,與2013年延平區(qū)20個(gè)豬場(chǎng)的豬偽狂犬病野毒感染調(diào)查結(jié)果56.26%相比,無(wú)下降,而從臨床上調(diào)查A、B兩場(chǎng)無(wú)爆發(fā)病情,說(shuō)明兩豬場(chǎng)豬群呈隱性感染,雖然2014年A、B兩場(chǎng)PR的得到了有效的控制,但隱形感染和種豬排毒現(xiàn)象較為嚴(yán)重,需制定有效的免疫程序,加強(qiáng)疫苗免疫,提高豬場(chǎng)的養(yǎng)殖條件。在本研究中,我們基于gE基因片段,建立的豬偽狂犬病PCR診斷方法具有良好的特異性,靈敏性和重復(fù)性。對(duì)野毒、疫苗毒以及陰性病料的檢測(cè)結(jié)果,與國(guó)標(biāo)一致。利用這一方法,我們抽檢了延平區(qū)12個(gè)豬場(chǎng)和A、B兩場(chǎng)的臍帶血和公豬精液。檢測(cè)結(jié)果如下：延平區(qū)12個(gè)豬場(chǎng)共抽檢132份臍帶血,PCR陽(yáng)性率為31.82%,A場(chǎng)陽(yáng)性率為57.14%,B場(chǎng)陽(yáng)性率為60.71%；12個(gè)場(chǎng)共抽檢公豬精液86份,陽(yáng)性率為15.12%,A、B兩場(chǎng)公豬精液PCR野毒陽(yáng)性率均為0.00%,由此推斷A、B兩豬場(chǎng)由母豬傳播給仔豬的垂直感染壓力大,高于延平區(qū)12個(gè)豬場(chǎng)的平均水平,而由公豬精液傳播的暫未造成危險(xiǎn)。由兩豬場(chǎng)的gE以及gB抗體檢測(cè)結(jié)果分析表明,兩個(gè)豬場(chǎng)豬群除公豬外都呈隱性感染,臨床無(wú)癥狀,可見(jiàn)豬群得到有效控制,生產(chǎn)較為穩(wěn)定,但由保育豬舍轉(zhuǎn)至肥育豬舍時(shí),gE抗體明顯升高,此階段仔豬母源抗體已消退,加上環(huán)境突變等因素,豬群由于應(yīng)激而容易被野毒感染,加上母豬排毒,使得病毒在場(chǎng)內(nèi)循環(huán),因此對(duì)仔豬首免及肥育豬監(jiān)測(cè)是穩(wěn)定gE抗體陽(yáng)性的關(guān)鍵。結(jié)論：雖然通過(guò)疫苗接種有效減少豬偽狂犬病的發(fā)生的散播,但大部分豬場(chǎng)并沒(méi)有徹底消滅豬偽狂犬病,部分免疫豬仍有排毒的可能,嚴(yán)重威脅易感豬。因此兩豬場(chǎng)需要對(duì)母豬進(jìn)行嚴(yán)格的免疫防控,盡可能淘汰陽(yáng)性豬以免給豬場(chǎng)造成生產(chǎn)繁殖壓力,保障新生仔豬的品質(zhì)。用ELISA檢測(cè)方法對(duì)豬場(chǎng)的不同豬群的PRV-gB抗體和PRV-gE抗體進(jìn)行血清學(xué)檢測(cè),并對(duì)其結(jié)果進(jìn)行評(píng)估,以作為判斷該豬場(chǎng)是否感染野毒、接種疫苗是否達(dá)到最佳效果的依據(jù),并結(jié)合PCR診斷方法分析評(píng)估被檢豬群的垂直傳播風(fēng)險(xiǎn),對(duì)指導(dǎo)規(guī)�；i場(chǎng)全面實(shí)施偽狂犬病凈化是有實(shí)際意義的。
[Abstract]:Objective: To evaluate and analyze the wild virus infection rate of porcine pseudorabies in two large-scale pig farms in Yanping District, Nanping, Fujian Province, and to establish a PCR diagnosis method to analyze the vertical transmission risk of wild poison in two pig pseudorabies by using the gE-ELISA and gB-ELISA kit. Method: two large-scale pig farms in Yanping District of Nanping city from November 2014 to December were entered. The gE antibody and gB antibody of the pig pseudorabies were detected and the results were statistically analyzed to evaluate the positive rate of wild virus infection in two large-scale pig farms. The statistical results were compared with the epidemiological survey results of the Nanping Yanping District in 2013 to evaluate the epidemic progress of the porcine pseudorabies virus (PRV) in the two pig farms in 2014. To establish a PCR diagnostic method for detection of wild poison in porcine pseudorabies (PR), and to analyze the specificity, sensitivity, repeatability and compliance with the national standard of the established PCR method. The PCR method established in this experiment was used to detect the umbilical cord blood and boar semen of the gE antibody positive pigs of two large-scale pig farms and 12 pig farms in Yanping District. The statistical comparison of two pigs was made. The PCR positive rate of field umbilical cord blood and semen and the average positive rate of 12 pig farms were used to assess the vertical transmission risk pressure in two large-scale pig farms. Results and analysis: a total of 353 serum samples were sampled from the A pig farm, and 267 serum samples were sampled from B piggery. The results of gE antibody test showed that all pigs in the pigs tested by A and B were examined in addition to the boars except for the boars. The positive rates of gE antibody were 51.08% and 58.80%, respectively. The positive rate of two fields was PR wild. The positive rate of gE antibody of two boars was 0%, which belonged to negative pigs. It showed that the boar group was not infected with PR and.A, and the positive rate of gE antibody of sow in B pig farm was 53.87%, 46.15%, and the positive rate of gE antibody in 30~40 day old pigs was divided. 26.09%, 51.16%, plus the proportion of suspicious numbers, respectively: 31.89%, 51.16%; the positive rates of gE antibody in 60 day old pigs were 45.45%, 65.96%, while the positive rate of gE antibody in the 90~120 days old pigs was up to 90%. Thus, it was concluded that the pig group was detected by PR wild venom.GB antibody at this stage when the transfer house was transferred to the fattening house. The results showed that the positive rate of gB antibody in two fields was 96.87%, 96.84%, and the positive rate of gB antibody in two pig farms was 100%, and the positive rate of gB antibody of sow was 93.4%, 100%, the level of antibody was high and the degree of dispersion was low. It may be a comprehensive cooperative use of vaccine immunization and PR wild virus infection, and the positive rate of gB antibody in 30-40 day old pigs. 94.20%, 97.67%, and high degree of uniformity, therefore, it can be concluded that in addition to the influence of mother antibody, the first immunization effect of two pig farms is good, and the antibody level A pig farm is higher than the B pig farm; the serum gB antibody S/N value of the two pig farm from 60 days to 120 days old decreases continuously, that is, the antibody level rises, and the maternal antibody has almost subsided, and the gE antibody also does not It was concluded that the average value of gE antibody positive rate in.2014 year A field of pig group at 60 days old, especially 120 days old, was 51.08%, and the average value of gE antibody positive rate of B field was 58.80%, compared with the result 56.26% of swine pseudorabies wild virus infection in 20 pig farms in Yanping area in 2013, no descent, but from clinical investigation A, B two fields were not exploded. It shows that the two pig farms are recessive infection in the two pig farm. Although the two fields of PR in 2014 and B have been effectively controlled, the stealth infection and the porcine detoxification are more serious. It is necessary to formulate effective immune procedures, strengthen the immunization of the vaccine and improve the breeding conditions of the pig farms. In this study, we based on the gE gene fragment to establish the PCR diagnosis of porcine pseudorabies. The method had good specificity, sensitivity and repeatability. The results of detection of wild, vaccine and negative diseases were the same as the national standard. Using this method, we checked the umbilical cord blood and boar semen of 12 pig farms and A and B fields in Yanping District. The results were as follows: 132 umbilical cord blood samples were sampled from 12 pig farms in Yanping District, and the positive rate of PCR was detected. For 31.82%, the positive rate of A field was 57.14%, the positive rate of B field was 60.71%, and 86 parts of the boar semen were detected in 12 fields, the positive rate was 15.12%, A, and the positive rate of PCR wild poison in the semen of the two boars was 0%. Thus, A was infer that the vertical infection pressure of the piglets transmitted by the sows in the B two pig was higher than the average level of the 12 pig farms in the Yanping District, and the semen passed from the boar semen. The analysis of the gE and gB antibody test results from two piggery showed that the two pig farms were infected with recessive infection except for the boars, and the clinical symptoms were asymptomatic. The pigs were effectively controlled and the production was more stable, but the anti body body of the piglets increased obviously when the pig houses were transferred to the fattening piggery, and the antibody of the mother source of the piglets had subsided at this stage. Because of the environmental mutation and so on, the pig group is easily infected by the wild virus because of stress, and the sow detoxification, causing the virus to circulate in the field. So the monitoring of the piglet first and the fattening pigs is the key to stabilizing the gE antibody positive. Conclusion: Although the vaccine inoculation effectively reduces the spread of porcine pseudorabies, most pigs are not thoroughly. To eliminate porcine pseudorabies, some pigs still have the possibility of detoxification, which seriously threaten the susceptible pigs. Therefore, the two piggery needs to be strictly immune and control the sows, to eliminate the positive pigs as much as possible so as to avoid the production pressure of the piggery and to ensure the quality of the newborn piglets. The PRV-gB antibody and PRV-gE of different pig farms in the pig farm are used by the ELISA test method. The antibody was tested by serology, and the results were evaluated as a basis for judging whether the pig farm was infected with wild poison and whether the vaccine reached the best effect, and the PCR diagnostic method was used to analyze the vertical transmission risk of the pigs tested. It was of practical significance to guide the purification of Pseudorabies in a large-scale pig farm.
【學(xué)位授予單位】：福建農(nóng)林大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2015
【分類(lèi)號(hào)】：S858.28

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本文編號(hào)：2058970