Agresti A, Ponti W, Rocchi M, Meneveri R, Marozzi A, Cavalleri D, Peri E, Poli G, Ginelli E
Department of Biology and Genetics for Medical Sciences, University of Milan, Italy.
Am J Vet Res. 1993 Mar;54(3):373-8.
A specific polymerase chain reaction (PCR) assay was devised, allowing detection of 1 bovine leukemia virus (BLV)-infected cell in 10(4) bovine lymphocytes. The efficacy of field application of the developed method was verified by evaluating the rate of viral transmission to calves from infected cows, whether they have persistent lymphocytosis. With this objective, 43 calves were simultaneously tested at birth and at 6 months of age for viral antibodies in serum and for proviral DNA in lymphocytes. At birth, 36 calves were BLV-negative and 3 were BLV-positive by results of serologic and DNA-based assays. Conversely, results for 4 calves had lack of correlation between the diagnostic methods. In particular, 2 calves were DNA-positive and antibody-negative for BLV and 2 other calves had the opposite test results. At 6 months of age, when the immunologic pattern more closely reflects the status of calves' immune response, independent of maternal antibodies, all calves DNA-negative for BLV at birth (n = 38), were consistently PCR- and antibody-negative for BLV. On the contrary, the cattle DNA-positive for BLV at birth (n = 5), whether seropositive or not, were PCR- and antibody-positive for BLV, at the time of the second screening. Thus, these results indicate reliability of the PCR to diagnose perinatal BLV infection. Furthermore, the observation that all calves found to be infected at birth were born to BLV-positive cows with persistent lymphocytosis, indicates that the persistent lymphocytosis status of the cow may represent a factor associated with BLV infection in utero.
设计了一种特异性聚合酶链反应(PCR)检测方法,能够在10⁴个牛淋巴细胞中检测出1个感染牛白血病病毒(BLV)的细胞。通过评估感染奶牛向犊牛的病毒传播率(无论这些奶牛是否患有持续性淋巴细胞增多症),验证了所开发方法在实际应用中的有效性。为此,在出生时和6月龄时,对43头犊牛同时进行血清中病毒抗体和淋巴细胞中前病毒DNA的检测。出生时,血清学检测和基于DNA的检测结果显示,36头犊牛BLV呈阴性,3头犊牛BLV呈阳性。相反,4头犊牛的诊断方法结果缺乏相关性。具体而言,2头犊牛BLV的DNA检测呈阳性但抗体检测呈阴性,另外2头犊牛的检测结果相反。在6月龄时,当免疫模式更能准确反映犊牛的免疫反应状态(不受母源抗体影响)时,所有出生时BLV的DNA检测呈阴性(n = 38)的犊牛,BLV的PCR检测和抗体检测均为阴性。相反,出生时BLV的DNA检测呈阳性(n = 5)的牛,无论血清学检测是否呈阳性,在第二次筛查时,BLV的PCR检测和抗体检测均为阳性。因此,这些结果表明PCR诊断围产期BLV感染具有可靠性。此外,所有出生时被发现感染的犊牛均出生于患有持续性淋巴细胞增多症的BLV阳性奶牛,这一观察结果表明,奶牛的持续性淋巴细胞增多症状态可能是子宫内BLV感染的相关因素。