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对用于诊断小反刍兽慢病毒感染的酶联免疫吸附测定(ELISA)策略组合的深入了解。

An insight into a combination of ELISA strategies to diagnose small ruminant lentivirus infections.

作者信息

de Andrés X, Ramírez H, Bertolotti L, San Román B, Glaria I, Crespo H, Jáuregui P, Minguijón E, Juste R, Leginagoikoa I, Pérez M, Luján L, Badiola J J, Polledo L, García-Marín J F, Riezu J I, Borrás-Cuesta F, de Andrés D, Rosati S, Reina R, Amorena B

机构信息

Institute of Agrobiotechnology (CSIC-UPNA-Gobierno de Navarra), Navarre, Spain.

出版信息

Vet Immunol Immunopathol. 2013 Apr 15;152(3-4):277-88. doi: 10.1016/j.vetimm.2012.12.017. Epub 2013 Jan 4.

Abstract

A single broadly reactive standard ELISA is commonly applied to control small ruminant lentivirus (SRLV) spread, but type specific ELISA strategies are gaining interest in areas with highly prevalent and heterogeneous SRLV infections. Short (15-residue) synthetic peptides (n=60) were designed in this study using deduced amino acid sequence profiles of SRLV circulating in sheep from North Central Spain and SRLV described previously. The corresponding ELISAs and two standard ELISAs were employed to analyze sera from sheep flocks either controlled or infected with different SRLV genotypes. Two outbreaks, showing SRLV-induced arthritis (genotype B2) and encephalitis (genotype A), were represented among the infected flocks. The ELISA results revealed that none of the assays detected all the infected animals in the global population analyzed, the assay performance varying according to the genetic type of the strain circulating in the area and the test antigen. Five of the six highly reactive (57-62%) single peptide ELISAs were further assessed, revealing that the ELISA based on peptide 98M (type A ENV-SU5, consensus from the neurological outbreak) detected positives in the majority of the type-A specific sera tested (Se: 86%; Sp: 98%) and not in the arthritic type B outbreak. ENV-TM ELISAs based on peptides 126M1 (Se: 82%; Sp: 95%) and 126M2 0,65 0.77 (Se: 68%; Sp: 88%) detected preferentially caprine arthritis encephalitis (CAEV, type B) and visna/maedi (VMV, type A) virus infections respectively, which may help to perform a preliminary CAEV vs. VMV-like typing of the flock. The use of particular peptide ELISAs and standard tests individually or combined may be useful in the different areas under study, to determine disease progression, diagnose/type infection and prevent its spread.

摘要

一种单一的具有广泛反应性的标准酶联免疫吸附测定(ELISA)通常用于控制小反刍兽疫慢病毒(SRLV)的传播,但在SRLV感染高度流行且异质性高的地区,类型特异性ELISA策略正受到关注。在本研究中,利用西班牙中北部绵羊中流行的SRLV和先前描述的SRLV的推导氨基酸序列图谱设计了短(15个残基)合成肽(n = 60)。采用相应的ELISA和两种标准ELISA分析来自受控或感染不同SRLV基因型的羊群的血清。在受感染的羊群中出现了两起疫情,分别表现为SRLV诱导的关节炎(B2基因型)和脑炎(A基因型)。ELISA结果显示,在分析的总体人群中,没有一种检测方法能检测到所有受感染动物,检测性能因该地区流行毒株的基因类型和检测抗原而异。对六种高反应性(57 - 62%)的单肽ELISA中的五种进行了进一步评估,结果显示基于肽98M(A基因型ENV-SU5,来自神经疫情的共识)的ELISA在大多数检测的A型特异性血清中检测到阳性(敏感性:86%;特异性:98%),而在关节炎B型疫情中未检测到阳性。基于肽126M1(敏感性:82%;特异性:95%)和126M2(敏感性:68%;特异性:88%)的ENV-TM ELISA分别优先检测到山羊关节炎脑炎病毒(CAEV,B型)和维斯纳/梅迪病毒(VMV,A型)感染,这可能有助于对羊群进行CAEV与VMV样的初步分型。单独或联合使用特定的肽ELISA和标准检测方法可能有助于在所研究的不同地区确定疾病进展、诊断/分型感染并预防其传播。

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