PDF(Pubmed)
Abstract：
UNASSIGNED: 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-gag (LTR-gag) region using samples from animals infected with subtypes of SRLV known to circulate in Poland.
UNASSIGNED: 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.
UNASSIGNED: 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.
UNASSIGNED: 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-gag region did prove to be, at least for genotypes A and B.
UNASSIGNED: 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.
UNASSIGNED: 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.
UNASSIGNED: 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-gag region did prove to be, at least for genotypes A and B.
摘要：
■小反刍动物慢病毒（SRLV）引起多系统，绵羊和山羊的变性和慢性疾病。有五种基因型(A，B,C,D和E),其中A和B是最普遍的。这项研究的目的是评估EradkitSRLV基因分型ELISA的血清分型效率和新开发的巢式实时PCR的分子分型效率，该PCR靶向长末端重复序列gag（LTR-gag）区域，使用感染了已知在波兰流通的SRLV亚型的动物样品。
■对34只绵羊和63只山羊的97份血清样本进行了免疫测定，用PCR检测了31只绵羊和55只山羊的86份DNA样本。所有反刍动物均被A1，A5，A12，A13，A16，A17，A18，A23，A24，A27，B1和B2亚型的已知SRLV菌株感染。
■总共69（80.2%，86个测试样品中的95％置信区间71.6％-88.8％）在PCR中给出了阳性结果。在86个样本中的17个(19.8%)中，未检测到SRLV的前病毒DNA。通过PCR进行的MVV（基因型A）和CAEV（基因型B）之间的区别总是与预先的系统发育分析相匹配。没有观察到交叉反应性。另一方面,通过较早的系统发育分析和EradikitSRLV基因分型ELISA进行基因分型的样本比例为42.3%.该测试无法对40.2%的样本进行分类，17.5%的血清分类不正确。
■我们的结果表明，EradikitSRLV基因分型试剂盒不是预测SRLV基因型的可靠方法，虽然基于LTR-gag区域的嵌套实时PCR确实被证明是，至少对于基因型A和B。
■对34只绵羊和63只山羊的97份血清样本进行了免疫测定，用PCR检测了31只绵羊和55只山羊的86份DNA样本。所有反刍动物均被A1，A5，A12，A13，A16，A17，A18，A23，A24，A27，B1和B2亚型的已知SRLV菌株感染。
■总共69（80.2%，86个测试样品中的95％置信区间71.6％-88.8％）在PCR中给出了阳性结果。在86个样本中的17个(19.8%)中，未检测到SRLV的前病毒DNA。通过PCR进行的MVV（基因型A）和CAEV（基因型B）之间的区别总是与预先的系统发育分析相匹配。没有观察到交叉反应性。另一方面,通过较早的系统发育分析和EradikitSRLV基因分型ELISA进行基因分型的样本比例为42.3%.该测试无法对40.2%的样本进行分类，17.5%的血清分类不正确。
■我们的结果表明，EradikitSRLV基因分型试剂盒不是预测SRLV基因型的可靠方法，虽然基于LTR-gag区域的嵌套实时PCR确实被证明是，至少对于基因型A和B。
