Geary S J
Bionique Laboratories, Inc., Saranac Lake, New York 12983.
Isr J Med Sci. 1987 Jun;23(6):747-51.
Mycoplasma gallisepticum causes respiratory diseases in the form of tracheitis and air sacculitis in chickens and turkeys. It is a major cause of reduced egg production, reduced hatchability, and downgrading of carcasses. Current means of diagnosis depend on the isolation and identification of the organisms, or on serological assays to detect serum antibodies. The evaluation of avian sera for M. gallisepticum antibodies is becoming more difficult to interpret, and thus less useful, due to the increasing use of killed M. gallisepticum vaccines. Maximum efficiency of M. gallisepticum disease control requires a rapid and sensitive identification system. A biotinylated total genome M. gallisepticum DNA probe was constructed by labeling the DNA with biotin-11-dUTP in a standard nick-translation reaction. Hybridization reactions with 100 ng/ml of biotinylated probe were capable of detecting 75 ng of M. gallisepticum target-DNA and 1.5 x 10(4) M. gallisepticum/ml within 24 h. The probe did hybridize to other mycoplasma DNAs, but to a greatly reduced degree.
鸡败血支原体可引发鸡和火鸡的气管炎症和气囊炎形式的呼吸道疾病。它是导致蛋产量下降、孵化率降低以及胴体等级降低的主要原因。目前的诊断方法依赖于病原体的分离与鉴定，或依赖血清学检测来检测血清抗体。由于鸡败血支原体灭活疫苗的使用日益增加，对禽血清中鸡败血支原体抗体的评估变得越来越难以解释，因而实用性降低。鸡败血支原体疾病控制的最大效率需要一个快速且灵敏的鉴定系统。通过在标准的缺口平移反应中用生物素-11-dUTP标记DNA，构建了一种生物素化的鸡败血支原体全基因组DNA探针。与100 ng/ml生物素化探针的杂交反应能够在24小时内检测到75 ng的鸡败血支原体靶DNA和每毫升1.5×10⁴个鸡败血支原体。该探针确实能与其他支原体DNA杂交，但杂交程度大大降低。
