引言猪肺炎支原体(Mycoplasma hyopneumoniae,Mhp)是引起猪支原体肺炎(Mycoplasma pneumoniae of swine,MPS)的主要病原,其致病机理为特异性的黏附于猪的呼吸道黏膜上皮组织,造成上皮细胞的病变或坏死,引发MPS的发生。本研究利用猪肺炎...
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引言猪肺炎支原体(Mycoplasma hyopneumoniae,Mhp)是引起猪支原体肺炎(Mycoplasma pneumoniae of swine,MPS)的主要病原,其致病机理为特异性的黏附于猪的呼吸道黏膜上皮组织,造成上皮细胞的病变或坏死,引发MPS的发生。本研究利用猪肺炎支原体强弱毒株体外感染猪呼吸道上皮细胞,从感染细胞的氧化损伤和差异表达蛋白两方面来分析猪肺炎支原体感染与宿主细胞应答过程中的相互关系。
引言牛副流感病毒3型（Bovine parainfluenza virus type 3,BPIV3）,为副粘病毒科（Paramyxoviridae）副粘病毒亚科（Paramyxovirinae）呼吸道病毒属（Respirovirus）的成员,是引起犊牛肺炎的主要病原之一。犊牛肺炎是一种临床常见的疾...
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引言牛副流感病毒3型（Bovine parainfluenza virus type 3,BPIV3）,为副粘病毒科（Paramyxoviridae）副粘病毒亚科（Paramyxovirinae）呼吸道病毒属（Respirovirus）的成员,是引起犊牛肺炎的主要病原之一。犊牛肺炎是一种临床常见的疾病,每年都给养牛业造成巨大的经济损失。BPIV3已在我国内蒙古、黑龙江、山东等地存在。HN蛋白和F蛋白是PIV3的保护性抗原,常用于亚单位疫苗的制备。本实
P65 protein, the major immunodominant protein of Mycoplasma hyopneu-moniae （Mhp） exhibiting no cross-reaction with other mycoplasmas, is general y used as a target protein for Mhp detection. In this study, BALB/c mi...
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P65 protein, the major immunodominant protein of Mycoplasma hyopneu-moniae （Mhp） exhibiting no cross-reaction with other mycoplasmas, is general y used as a target protein for Mhp detection. In this study, BALB/c mice were immunized with prokaryotical y expressed P65 recombinant protein to prepare monoclonal anti-body. After screening with Mhp whole-cel protein and P65 protein, a specific hy-bridoma cel line, 3G12, was obtained by ELISA. Identification results indicated that the antibody secreted by 3G12 hybridoma cel s could react with P65 protein and Mhp whole-cel protein. According to indirect ELISA assay, 3G12 cel culture super-natant possessed a titer of 1∶12 800 against P65 protein and 1∶3 200 against Mhp whole-cel protein; 3G12 ascites possessed a titer of above 1∶4 000 000 against P65 protein and above 1∶20 000 against Mhp 168 whole-cel protein. After long-term in vitro culture and continuous freezing-thawing, 3G12 cel line could stably secrete antibodies. A monoclonal antibody against P65 protein and Mhp whole-cel protein was successful y obtained in the present study, which provided basis for further in-vestigating the pathogenic mechanism of Mhp and establishing diagnostic methods of Mycoplasmal pneumonia of swine （MPS）.
[Objective] 303 nasal swabs samples were collected from pigs in farms located in Taizhou city, Jiangsu Province, China from March to December 2012 for the purpose of detecting the presence of Mycoplasma hyopneumoniae,...
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[Objective] 303 nasal swabs samples were collected from pigs in farms located in Taizhou city, Jiangsu Province, China from March to December 2012 for the purpose of detecting the presence of Mycoplasma hyopneumoniae, the primary agent of Enzootic porcine pneumonia （EPP） in pig herds using the nested PCR and Real time PCR techniques. [Method] Nasal swabs were collected from pigs of different ages＇ i.e. 7, 14, 21, 28, 30 and 35 days old, soaked in sterile 1 xPBS overnight at 4 ℃ and DNA extracted using the TIANamp（R） bacterial DNA kit. The DNA samples underwent amplification under the Mhyo 183 q-PCR and P36 primer Nested PCR systems. [Result] With the Nested PCR assay, 38 （12.5%） out of 303 samples tested positive for the presence of M. hyopneumoniae; with the real time PCR assay 152 （50.2%） tested positive for M. hyopneumoniae. The two assays matched to positively detect Mhyo in 22 （7.3%） samples and again matched in 127 （41.9%） samples negative for Mhyo infection. The pattern of infection in both assays was similar where 7- and 35-day-old piglets in both assays had the highest rates of infection i.e. 15.6% and 18.4% for n-PCR and 53.1% and 56.6% for q-PCR for 7- and 35-day-old piglets respectively. [Conclusion] The results highlight the suitability of both PCR assays in establishing the herd infection status of pigs in field conditions.
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