Comparative Analysis for the Detection and Monitoring of Mycoplasma hyopneumoniae Infection by Nested PCR(n-PCR) and Real time PCR(q-PCR) from Field Swine Herds

作     者：William Keeru KIMARU 白方方 武昱孜 Joyce Wanjiru MAINGI 华利忠 刘茂军 张旭 邵国青 鲍恩东 

作者机构：南京农业大学动物医学院江苏南京210014 江苏省农业科学院兽医研究所/农业部兽用生物制品工程技术重点实验室/国家兽用生物制品工程技术研究中心江苏南京210014 

出 版 物：《Agricultural Science & Technology》 (农业科学与技术（英文版）)

年 卷 期：2014年第15卷第6期

页      面：918-921页

学科分类：090603[农学-临床兽医学] 09[农学] 0906[农学-兽医学] 

基　　金：Supported by the Agricultural Science Independent Innovation Foundation of Jiangsu Province[CX(12)1001] 

主　　题：Enzootic porcine pneumonia Infection Mycoplasma hyopneumoniae PCR 

摘      要：[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.