利用 PCR 诊断技术评估达吉斯坦共和国牛白血病病毒 (BLV) 的发病率

D. A. Baboshko, K. A. Elfimov, M. G. Daudova, Kh. G. Koychuev, Kh. F.‐K. Gapizova, N. M. Gashnikova
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引用次数: 0

摘要

目的利用 PCR 诊断技术评估达吉斯坦共和国牛群中白血病病毒的发病率,并研究循环病毒的分子遗传特征。 材料和方法检测了 150 份牛血样。使用 RealBest-Vet DNA BLV 检测系统和一套实验室引物对样本进行了牛白血病病毒(BLV)的 PCR 诊断。使用 Sanger 方法对部分样本进行了测序,并对其进行了系统发育分析。 结果在 150 份样本中,有 24 份样本的 BLV 检测结果呈阳性。在 Untsukulskiy 地区,16 份样本中均未检测到 BLV DNA。在卡拉布达赫肯茨基(Karabudakhkentskiy)区的 40 个样本中,有 2 个样本(5%)检测到 BLV;在布伊纳茨基区(Buynakskiy)的 30 个样本中,有 1 个样本(3.3%)检测到 BLV;在巴巴伊托夫斯基(Babayurtovskiy)区的 60 个样本中,有 21 个样本(35%)检测到 BLV。在 13 个 BLV 阳性样本中,获得并破译了 1000 bp 的 env 基因片段。根据系统发育分析,7 个 BLV 样本属于 BLV 的第 7 个基因型,6 个属于 BLV 的第 4 个基因型。在巴巴耶尔托夫斯基地区分离到的 BLV 基因型 4 与哈萨克斯坦的病毒聚类,而其他农场的基因型 4 病毒则与俄罗斯 BLV 聚类。所研究的基因型 4 样本没有形成共同的聚类。相反,在巴巴耶尔托夫斯基地区农场分离到的 BLV 基因 7 型病毒,其序列组合成了一个基因相似的病毒群。 结论达吉斯坦农场牲畜白血病病毒发病率存在显著差异。在牛的病例登记中,没有发现与特定品种的牛或动物年龄有关的模式。研究表明,7 型和 4 型基因型病毒都在共和国境内流行。就 BLV 4 而言,可以推测其通过不同途径进入农场,但没有发现相关的病毒传播链。至于 BLV 基因型 7,已发现 BLV 的传播,这表明有必要加强农场的白血病预防措施。
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Assessment of the incidence of leukemia virus (BLV) in cattle in the Republic of Dagestan using PCR diagnostics
   Aim. Assessment of the incidence of leukemia virus in cattle using PCR diagnostics in herds of the Republic of Dagestan and study of the molecular genetic characteristics of circulating viruses.   Materials and Methods. 150 cattle blood samples were examined. PCR diagnostics of samples for the presence of bovine leukosis virus (BLV) were carried out using the RealBest‐Vet DNA BLV test system and a laboratory set of primers. Some of the samples were sequenced using the Sanger method and their phylogenetic analysis was performed.   Results. Out of 150 samples, 24 samples were positive for the presence of BLV. In the Untsukulskiy district, no BLV DNA was detected in any of the 16 samples. In the Karabudakhkentskiy district out of 40 – in 2 (5 %), in Buynakskiy district – in 1 out of 30 (3.3 %) and in Babayurtovskiy district–in 21 out of 60 samples BLV was detected (35 %). For 13 BLV‐positive samples, fragments of the env gene measuring 1000 bp were obtained and deciphered. According to phylogenetic analysis, 7 samples of BLV belong to the 7th, and 6 – to the 4th genotype of BLV. The BLV genotype 4 isolated in the Babayurtovskiy district clusters with viruses from Kazakhstan, while viruses of genotype 4 from other farms cluster with Russian BLV. The studied samples of genotype 4 did not form common clusters. For the BLV genotype 7 isolated in farms of the Babayurtovskiy district, on the contrary, a combination of sequences into one cluster of genetically similar viruses was observed.   Conclusion. Significant differences in the incidence of leukemia virus in livestock on farms in Dagestan were revealed. No patterns were found in the registration of cattle cases with a specific breed of cattle or with the age of the animal. It has been shown that viruses of both genotypes 7 and 4 circulate in the Republic. For BLV 4, it is assumed that there are different ways of its introduction into farms but no associated chains of virus spread have been found. For BLV genotype 7, transmission of BLV has been registered, which indicates the need to strengthen leukemia prevention measures on farms.
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