Kasten R W, Carpenter T E, Snipes K P, Hirsh D C
Department of Pathology, Microbiology, and Immunology, School of Veterinary Medicine, University of California, Davis 95616, USA.
Avian Dis. 1997 Jul-Sep;41(3):676-82.
A polymerase chain reaction (PCR)-based assay using primers constructed to amplify the gene (psl) encoding the P6-like protein (Psl) of Pasteurella multocida was developed. After Southern blotting and hybridization with psl, the assay (PCR-H) was found to be specific (it did not detect a variety of other avian bacterial pathogens) and sensitive (detected > or = 10 P. multocida organisms or > or = 24 femtograms of extracted P. multocida DNA). Samples were collected from the oropharynx of randomly selected birds housed on premises that had recently experienced an outbreak of avian cholera (outbreak farms) or from birds housed on premises that had not reported an outbreak of this disease during the preceding 12 mo (control farms). The PCR-H assay detected 11 infected turkeys out of a total of 178 sampled on six outbreak farms as compared with isolation of P. multocida from 23 turkeys by using mouse inoculation. Neither method detected P. multocida in samples collected from 174 turkeys sampled on six control farms. Statistical analysis using the Kappa test demonstrated that the results of the two tests showed poor agreement from five outbreak flocks (K = 0, 0, 0, 0.35, 0.47) and strong agreement from one outbreak flock (K = 0.89). Combined results from the outbreak flocks showed poor agreement (K = 0.49) between the two methods.
开发了一种基于聚合酶链反应（PCR）的检测方法，该方法使用构建的引物来扩增编码多杀性巴氏杆菌P6样蛋白（Psl）的基因（psl）。在进行Southern印迹并用psl进行杂交后，发现该检测方法（PCR-H）具有特异性（未检测到多种其他禽源细菌病原体）和敏感性（可检测到≥10个多杀性巴氏杆菌菌体或≥24飞克提取的多杀性巴氏杆菌DNA）。样本取自随机选择的鸟类的口咽，这些鸟类饲养在最近发生禽霍乱疫情的场所（疫情爆发养殖场），或取自在前12个月内未报告过该疾病疫情的场所饲养的鸟类（对照养殖场）。在六个疫情爆发养殖场采集的178只样本火鸡中，PCR-H检测方法检测到11只感染火鸡，而通过小鼠接种从23只火鸡中分离出多杀性巴氏杆菌。两种方法在从六个对照养殖场采集的174只火鸡样本中均未检测到多杀性巴氏杆菌。使用Kappa检验进行统计分析表明，两种检测方法的结果在五个疫情爆发鸡群中一致性较差（K = 0、0、0、0.35、0.47），在一个疫情爆发鸡群中一致性较强（K = 0.89）。疫情爆发鸡群的综合结果显示两种方法之间一致性较差（K = 0.49）。
