[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,...[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.展开更多
基金Supported by the Agricultural Science Independent Innovation Foundation of Jiangsu Province[CX(12)1001]~~
文摘[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.
