Tuyển tập các báo cáo nghiên cứu về y học được đăng trên tạp chí y học Respiratory Research cung cấp cho các bạn kiến thức về ngành y đề tài: BLV-CoCoMo-qPCR: Quantitation of bovine leukemia virus proviral load using the CoCoMo . | Jimba et al. Retrovirology 2010 7 91 http content 7 1 91 RETR0VIR0L0GY RESEARCH Open Access BLV-CoCoMo-qPCR Quantitation of bovine leukemia virus proviral load using the CoCoMo algorithm 1 3 4 Mayuko Jimba 1 Shin-nosuke Takeshima Kazuhiro Matoba Daiji Endoh Yoko Aida 1 Abstract Background Bovine leukemia virus BLV is closely related to human T-cell leukemia virus HTLV and is the etiological agent of enzootic bovine leukosis a disease characterized by a highly extended course that often involves persistent lymphocytosis and culminates in B-cell lymphomas. BLV provirus remains integrated in cellular genomes even in the absence of detectable BLV antibodies. Therefore to understand the mechanism of BLV-induced leukemogenesis and carry out the selection of BLV-infected animals a detailed evaluation of changes in proviral load throughout the course of disease in BLV-infected cattle is required. The aim of this study was to develop a new quantitative real-time polymerase chain reaction PCR method using Coordination of Common Motifs CoCoMo primers to measure the proviral load of known and novel BLV variants in clinical animals. Results Degenerate primers were designed from 52 individual BLV long terminal repeat LTR sequences identified from 356 BLV sequences in GenBank using the CoCoMo algorithm which has been developed specifically for the detection of multiple virus species. Among 72 primer sets from 49 candidate primers the most specific primer set was selected for detection of BLV LTR by melting curve analysis after real-time PCR amplification. An internal BLV TaqMan probe was used to enhance the specificity and sensitivity of the assay and a parallel amplification of a single-copy host gene the bovine leukocyte antigen DRA gene was used to normalize genomic DNA. The assay is highly specific sensitive quantitative and reproducible and was able to detect BLV in a number of samples that were negative using the previously developed nested PCR
