本文關(guān)鍵詞：高致病性豬繁殖與呼吸綜合征病毒TJM-F92株減毒活疫苗臨床免疫效果評價與分析　出處：《中國農(nóng)業(yè)大學(xué)》2016年博士論文　論文類型：學(xué)位論文

  更多相關(guān)文章： 豬繁殖與呼吸綜合征 減毒活疫苗 安全性 免疫效果 臨床評價

【摘要】：我國普遍采用減毒活疫苗免疫控制豬繁殖與呼吸綜合征病毒(PRRSV)感染,但減毒活疫苗的臨床安全性與效力受到關(guān)注。本研究以高致病性豬繁殖與呼吸綜合征病毒TJM-F92株減毒活疫苗為對象,開展了其臨床應(yīng)用情況、安全性及免疫效果的評價與分析,以期為養(yǎng)豬實際生產(chǎn)中合理使用TJM-F92株減毒活疫苗提供科學(xué)依據(jù)。采用調(diào)查問卷的方式,2015年3月至5月對24個省的720個豬場進行了TJM-F92株減毒活疫苗應(yīng)用情況的調(diào)查。結(jié)果顯示,收到反饋信息的700個豬場中,母豬群使用TJM-F92株減毒活疫苗的安全性好、較好和一般的豬場分別占52.29%、40.57%和7.14%,93.13%的豬場采用對母豬群每年普免3-4次的方式；在生長豬群使用TJM-F92株減毒活疫苗的634個豬場中,免疫效果好、較好和不明顯的豬場分別占29.96%、59.31%和10.73%。此外,分析發(fā)現(xiàn)生長豬的免疫效果與豬場位置、規(guī)模、生產(chǎn)方式以及采用的免疫程序有關(guān)。為進一步評價感染不同PRRSV毒株的豬場使用TJM-F92株減毒活疫苗的免疫效果,分析了我國豬群感染PRRSV毒株的狀況和特點。2014年4月至2016年4月,對采集自15個省56個豬場的經(jīng)RT-PCR檢測PRRSV ORF7核酸為陽性的163份樣本進行了ORF3、ORF5和NSP2高變區(qū)基因的擴增與序列測定和分析。結(jié)果顯示,擴增的130個ORF3基因、139個ORF5基因和113個NSP2基因片段均屬于PRRSV基因2型毒株,均可分為5個亞群,其中ORF3和ORF5基因各存在一個新亞群；在擴增的基因中,與高致病性PRRSV (HP-PRRSV)高度同源的ORF3、 ORF5、NSP2基因分別為68.46%、55.4%和84.07%;NSP2呈現(xiàn)11種缺失模式。通過對各個豬場、各個樣本的3個目標(biāo)基因的綜合分析揭示了豬場臨床感染PRRSV毒株的復(fù)雜性,研究共發(fā)現(xiàn)9類PRRSV野毒株,18類PRRSV毒株感染類型。HP-PRRSV、類NADC30、類VR-2332在豬群中出現(xiàn)的頻率分別為66.07%、26.79%、8.93%。單獨感染HP-PRRSV、類NADC30、類VR-2332等3類典型毒株的感染類型的豬場比例為53.57%,其中,單獨感染HP-PRRSV毒株的豬場占比最高,為41.07%。感染疑似復(fù)雜毒株和2類或2類以上毒株混合感染類型的豬場比例為45.64%,其中,HP-PRRSV和類NADC30混合感染的毒株感染類型的豬場占比最高,為12.5%。為分析TJM-F92株減毒活疫苗在不同PRRSV毒株感染類型豬群的臨床效果,對PRRSV毒株感染情況清楚的35個豬場,進行了TJM-F92株減毒活疫苗免疫后的臨床效果調(diào)查。結(jié)果顯示：免疫TJM-F92株減毒活疫苗后,臨床改善顯著、有改善、無改善的豬場比例分別為13/35、13/35、9/35。進一步的分析揭示了TJM-F92株減毒活疫苗在不同PRRSV毒株感染類型豬場的效果差異。單獨感染HP-PRRSV毒株的豬場,免疫后改善顯著、有改善和無改善的比例分別為50%、38.5%和12.5%,而類NADC30毒株感染豬場,疫苗免疫后未見顯著改善。為進一步分析豬群持續(xù)TJM-F92株減毒活疫苗免疫后的臨床安全性和效果,2014年7月至2016年1月追蹤了一個PRRSV感染豬場使用疫苗前后的母豬產(chǎn)仔性能、分娩率和保育豬臨床癥狀、各階段成活率的情況,并定期采用ELISA和基因擴增與序列測定分別對免疫后的生長豬群PRRSV抗體變化和保育豬群的PRRSV毒株感染的情況進行了監(jiān)測。結(jié)果顯示,母豬群免疫TJM-F92株減毒活疫苗前后,其產(chǎn)仔性能、分娩率、斷奶豬成活率差異都不顯著。母豬在不同妊娠階段接種TJM-F92株疫苗,對其產(chǎn)仔性能有不同的影響,在妊娠0-30大期間接種,相比其它妊娠階段接種,母豬窩均木乃伊數(shù)最高,且差異顯著,提示TJM-F92株減毒活疫苗在此妊娠階段免疫存在不安全性。生長豬群免疫后,其臨床癥狀顯著改善,保育期與育肥期成活率分別為96.97±1.48%、97.50±0.61%,與疫苗免疫前的差異極顯著。此外,發(fā)現(xiàn)保育豬的成活率與PRRSV感染時間以及毒株有關(guān)。由此表明,長期免疫TJM-F92株減毒活疫苗,有助于降低HP-PRRSV野毒株在豬群中的循環(huán)。綜上所述,我們的研究較為系統(tǒng)地評價了TJM-F92株減毒活疫苗的臨床安全性和免疫效果,對養(yǎng)豬生產(chǎn)中該疫苗的合理應(yīng)用有指導(dǎo)價值。
[Abstract]:The attenuated live vaccine is widely used to control the infection of porcine reproductive and respiratory syndrome virus (PRRSV) in China. However, the clinical safety and efficacy of attenuated live vaccine is concerned. In this study, highly pathogenic porcine reproductive and respiratory syndrome virus TJM-F92 strain live attenuated vaccine for the object, to carry out the analysis and evaluation of its clinical application, safety and immune effect, in order to raise the actual production and rational use of TJM-F92 strain live attenuated vaccine to provide the scientific basis for. Questionnaires were used to investigate the application of TJM-F92 live attenuated vaccine to 720 pig farms in 24 provinces from March 2015 to May. The results showed that 700 pig farms receive feedback information, sow group TJM-F92 strain live attenuated vaccine safety is good, good and the general farms accounted for 52.29%, 40.57% and 7.14%, 93.13% pig farms use of sows group every year tax-free 3-4 times; in the growth of pigs using live attenuated vaccine strain TJM-F92 the 634 pig farms, the immune effect is good, good and not obvious farms accounted for 29.96%, 59.31% and 10.73%. In addition, it is found that the immune effect of growing pigs is related to the location, size, mode of production and the immune procedures used. In order to further evaluate the immunization effect of live attenuated strain TJM-F92 of swine infected with different PRRSV strains, the status and characteristics of swine infected PRRSV strains in China were analyzed. From April 2014 to April 2016, we amplified and sequenced and analyzed the ORF3, ORF5 and NSP2 hypervariable regions in 163 samples collected from 56 pig farms in 15 provinces and tested positive for PRRSV ORF7 nucleic acid by RT-PCR. The results showed that 130 ORF3 genes, 139 ORF5 genes and 113 NSP2 genes amplified fragments belong to the PRRSV genotype 2 strains can be divided into 5 subgroups, including ORF3 and ORF5 gene are a subset of; in the amplified gene, and highly pathogenic PRRSV (HP-PRRSV) high degree ORF3, ORF5, NSP2 homologous genes were 68.46%, 55.4% and 84.07%; NSP2 showed 11 missing pattern. Through the comprehensive analysis of 3 target genes in pig farms and samples, the complexity of PRRSV infection in swine farms was revealed. 9 types of PRRSV wild strains and 18 kinds of PRRSV strains were found. The frequencies of HP-PRRSV, NADC30 and VR-2332 in the pigs were 66.07%, 26.79% and 8.93% respectively. The infection rate of 3 typical strains infected with HP-PRRSV, NADC30 and VR-2332 was 53.57%, and the proportion of pigs infected with HP-PRRSV strain was 41.07%. The proportion of mixed farms infected with suspected complex strains and 2 or more than 2 kinds of strains was 45.64%, of which HP-PRRSV and NADC30 mixed infection strains accounted for the highest proportion of infected pig farms, 12.5%. In order to analyze the clinical effect of attenuated live vaccine of TJM-F92 strain in different PRRSV infected pigs, the clinical effect of live attenuated TJM-F92 strain vaccine was investigated in 35 pig farms with clear infection of PRRSV. The results showed that after immunized TJM-F92 strain live attenuated vaccine, the proportion of clinical improvement, improvement and no improvement were 13/35, 13/35 and 9/35, respectively. Further analysis revealed the difference in the effect of TJM-F92 strain live attenuated vaccine in different type of swine farms with different PRRSV strains. The proportion of pigs that were infected with HP-PRRSV virus alone significantly improved, improved and no improvement was 50%, 38.5% and 12.5%, respectively, while NADC30 like strains were not significantly improved after vaccination. For further analysis of swine TJM-F92 strain for live attenuated vaccine immunity after the clinical safety and efficacy, from July 2014 to January 2016 followed a PRRSV infection using pig vaccine before and after the survival rate of sows farrowing performance, delivery rate and conservation in various stages of clinical symptoms in pigs, the situation, and regularly by ELISA and gene amplification was monitored with the sequencing respectively. After immunization the growth of pig PRRSV antibody changes and conservation herd strains of PRRSV infection. The results showed that the baby performance, delivery rate and the survival rate of weanling pigs were not significant before and after the sow group immunized with TJM-F92 attenuated live vaccine. In the different stage of gestation sows inoculated with TJM-F92 vaccine, have different effects on the reproductive performance, vaccination during pregnancy 0-30, compared to the other stage of pregnancy sows were vaccinated, the highest number of mummy nest, and the difference was significant, suggesting that TJM-F92 attenuated live vaccine on the immune stage of pregnancy have security. After growing pigs, their clinical symptoms were significantly improved. The survival rate was 96.97 + 1.48% and 97.50 97.50, respectively. In addition, the survival rate of the protected pigs was related to the time of PRRSV infection and the strain. Therefore, the long-term immune TJM-F92 strain attenuated live vaccine can help to reduce the circulation of HP-PRRSV wild strains in the pigs. To sum up, our research systematically evaluated the clinical safety and immune effects of live attenuated vaccine of TJM-F92 strain, which is of guiding value to the rational application of the vaccine in pig production.
【學(xué)位授予單位】：中國農(nóng)業(yè)大學(xué)
【學(xué)位級別】：博士
【學(xué)位授予年份】：2016
【分類號】：S858.28
，

本文編號：1346312