Koenig Patricia, Hoffmann Bernd, Depner Klaus R, Reimann Ilona, Teifke Jens P, Beer Martin
Institute of Diagnostic Virology, Friedrich-Loeffler-Institut, Boddenblick 5a, 17493 Greifswald-Insel Riems, Germany.
Vet Microbiol. 2007 Mar 10;120(3-4):343-51. doi: 10.1016/j.vetmic.2006.10.034. Epub 2006 Dec 4.
Attenuated live classical swine fever (CSF) viruses are the most efficacious vaccines against the disease. However, little is known about the distribution and detection of CSF vaccine viruses in the host. We therefore compared the new recombinant attenuated marker vaccine virus CP7_E2alf with the conventional C-strain vaccine concerning virus isolation, antigen-, and genome-detection in different samples within the first 42 days post-vaccination (p.v.). Leukocytes and several organs such as tonsils, lymph nodes, spleen, thymus, parotis and kidney were also tested using highly sensitive real-time reverse transcription-polymerase chain reaction (RT-PCR) techniques. It was demonstrated that vaccine virus could be detected by live animal sampling only in a few leukocytes samples at very low titres and genome copy numbers within the first 14 days after immunisation. Vaccine virus could also be isolated from individual tonsil samples within the first 6 days after vaccine application. In contrast, vaccine virus genomes were consistently detected in the tonsils up to day 42 by real-time RT-PCR. Distribution, amount of virus and viral genome levels were similar for both tested vaccines. In conclusion, blood samples could be the sample material of choice for detecting CSF wild type virus infection even in vaccinated animals after more than 14 days p.v., while tonsil sampling provided appropriate material for long-term detection of both tested CSF vaccine viruses using real-time RT-PCR methods.
减毒活经典猪瘟(CSF)病毒是预防该疾病最有效的疫苗。然而,关于CSF疫苗病毒在宿主体内的分布和检测情况却知之甚少。因此,我们比较了新型重组减毒标记疫苗病毒CP7_E2alf与传统C株疫苗在接种后(p.v.)前42天内不同样本中的病毒分离、抗原和基因组检测情况。还使用高灵敏度实时逆转录-聚合酶链反应(RT-PCR)技术对白细胞以及扁桃体、淋巴结、脾脏、胸腺、腮腺和肾脏等多个器官进行了检测。结果表明,免疫后前14天内,仅在少数白细胞样本中能以极低的滴度和基因组拷贝数通过活体动物采样检测到疫苗病毒。在接种疫苗后的前6天内,也能从单个扁桃体样本中分离出疫苗病毒。相比之下,通过实时RT-PCR在第42天前的扁桃体中始终能检测到疫苗病毒基因组。两种受试疫苗的病毒分布、病毒量和病毒基因组水平相似。总之,即使在接种后超过14天的动物中,血样也可能是检测CSF野生型病毒感染的首选样本材料,而扁桃体采样为使用实时RT-PCR方法长期检测两种受试CSF疫苗病毒提供了合适的材料。