Olech Monika, Kuźmak Jacek
Department of Pathology, Pulawy, Poland.
Department of Biochemistry, National Veterinary Research Institute, 24-100 Pulawy, Poland.
J Vet Res. 2024 Apr 30;68(2):181-188. doi: 10.2478/jvetres-2024-0025. eCollection 2024 Jun.
Small ruminant lentiviruses (SRLV) cause multisystemic, degenerative and chronic disease in sheep and goats. There are five genotypes (A, B, C, D and E), of which A and B are the most widespread. The purpose of this study was to evaluate the serotyping efficiency of the Eradikit SRLV Genotyping ELISA and the molecular typing efficiency of a newly developed nested real-time PCR targeting the long terminal repeat- (LTR-gag) region using samples from animals infected with subtypes of SRLV known to circulate in Poland.
A total of 97 sera samples taken from 34 sheep and 63 goats were immunoassayed, and 86 DNA samples from 31 sheep and 55 goats were tested with the PCR. All ruminants were infected with known SRLV strains of the A1, A5, A12, A13, A16, A17, A18, A23, A24, A27, B1 and B2 subtypes.
A total of 69 (80.2%, 95% confidence interval 71.6%-88.8%) out of 86 tested samples gave positive results in the PCR. In 17 out of the 86 (19.8%) samples, no proviral DNA of SRLV was detected. The differentiation between MVV (genotype A) and CAEV (genotype B) by PCR matched the predating phylogenetic analysis invariably. No cross-reactivity was observed. On the other hand, the proportion of samples genotyped the same by the older phylogenetic analysis and the Eradikit SRLV Genotyping ELISA was 42.3%. The test was unable to classify 40.2% of samples, and 17.5% of sera were incorrectly classified.
Our results showed that the Eradikit SRLV genotyping kit is not a reliable method for predicting SRLV genotype, while the nested real-time PCR based on the LTR- region did prove to be, at least for genotypes A and B.
小反刍兽疫病毒（SRLV）可导致绵羊和山羊出现多系统、退行性和慢性疾病。该病毒有五种基因型（A、B、C、D和E），其中A和B最为常见。本研究旨在利用波兰已知流行的SRLV亚型感染动物的样本，评估Eradikit SRLV基因分型酶联免疫吸附测定（ELISA）的血清分型效率以及针对长末端重复序列-（LTR-gag）区域新开发的巢式实时聚合酶链反应（PCR）的分子分型效率。
对从34只绵羊和63只山羊采集的97份血清样本进行免疫分析，并用PCR检测从31只绵羊和55只山羊采集的86份DNA样本。所有反刍动物均感染了已知的A1、A5、A12、A13、A16、A17、A18、A23、A24、A27、B1和B2亚型的SRLV毒株。
86份检测样本中，共有69份（80.2%，95%置信区间71.6%-88.8%）在PCR检测中呈阳性结果。86份样本中有17份（19.8%）未检测到SRLV的前病毒DNA。通过PCR区分梅迪-维斯纳病毒（MVV，基因型A）和山羊关节炎脑炎病毒（CAEV，基因型B）与之前的系统发育分析结果始终一致。未观察到交叉反应。另一方面，通过较旧的系统发育分析和Eradikit SRLV基因分型ELISA对相同样本进行基因分型的比例为42.3%。该检测无法对40.2%的样本进行分类，17.5%的血清被错误分类。
我们的结果表明，Eradikit SRLV基因分型试剂盒不是预测SRLV基因型的可靠方法，而基于LTR区域的巢式实时PCR至少对于基因型A和B来说是可靠的。
