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评估三种用于检测驯鹿(Rangifer tarandus tarandus)抗α疱疹病毒抗体的商用牛酶联免疫吸附测定试剂盒。

Evaluation of three commercial bovine ELISA kits for detection of antibodies against Alphaherpesviruses in reindeer (Rangifer tarandus tarandus).

作者信息

Das Neves Carlos G, Roger Matthieu, Yoccoz Nigel G, Rimstad Espen, Tryland Morten

机构信息

The Norwegian School of Veterinary Science, Department of Food Safety and Infection Biology, Section of Arctic Veterinary Medicine, Stakkevollveien 23, NO-9010 Tromsø, Norway.

出版信息

Acta Vet Scand. 2009 Mar 9;51(1):9. doi: 10.1186/1751-0147-51-9.

Abstract

BACKGROUND

The genus Varicellovirus (family Herpesviridae subfamily Alphaherpesvirinae) includes a group of viruses genetically and antigenically related to bovine herpesvirus 1 (BoHV-1) among which cervid herpesvirus 2 (CvHV-2) can be of importance in reindeer. These viruses are known to be responsible for different diseases in both wild and domestic animals. Reindeer are a keystone in the indigenous Saami culture and previous studies have reported the presence of antibodies against alphaherpesviruses in semi-domesticated reindeer in northern Norway. Mortality rates, especially in calves, can be very high in some herds and the abortion potential of alphaherpesvirus in reindeer, unlike in bovines, remains unknown. ELISA kits are the most used screening method in domestic ruminants and given the close genetic relationship between viruses within this genus, it might be possible to use such kits to screen cervids for different alphaherpesviruses. We have compared three different commercial ELISA kits in order to validate its use for reindeer and CvHV-2.

METHODS

Three commercial bovine ELISA kits (A, B and C), using either indirect (A) or blocking (B and C) ELISA techniques to detect antibodies against BoHV-1 were tested with sera from 154 reindeer in order to detect antibodies against CvHV-2. A Spearman's rank-based coefficient of correlation (rho) was calculated. A dilution trial was performed for all kits. A virus neutralization test using both BoHV-1 and CvHV-2 was carried out.

RESULTS

Seroprevalence was almost the same with all kits (40-41%). Despite a similar qualitative score, quantitatively kits classified samples differently and a strong correlation was only identified between Kits B and C. Blocking kits performed better in both repeatability and in the dilution trial. The virus neutralization results confirmed the ELISA results to a very high degree. Neutralizing titres ranged from 1:2 to 1:256 and from 0 to 1:16 against CvHV-2 and BoHV-1 respectively.

CONCLUSION

Results show that the genetic and antigenic similarity between BoHV-1 and CvHV-2 enables the use of a bovine gB blocking ELISA kit to screen reindeer. The use of an ELISA kit is both cheaper and time saving, allowing screening of large populations. This study revealed a high number of positive animals against CvHV-2 and its impact and distribution in the general population should be further evaluated.

摘要

背景

水痘病毒属(疱疹病毒科甲型疱疹病毒亚科)包括一组在基因和抗原方面与牛疱疹病毒1(BoHV-1)相关的病毒,其中鹿疱疹病毒2(CvHV-2)可能对驯鹿具有重要意义。已知这些病毒会导致野生动物和家畜患上不同疾病。驯鹿是萨米族本土文化的关键元素,此前的研究报告称,挪威北部半驯化驯鹿体内存在抗甲型疱疹病毒的抗体。在一些鹿群中,死亡率,尤其是幼鹿的死亡率可能非常高,而且与牛不同,驯鹿中甲型疱疹病毒的流产风险仍然未知。酶联免疫吸附测定(ELISA)试剂盒是家畜反刍动物中最常用的筛查方法,鉴于该属内病毒之间密切的基因关系,有可能使用此类试剂盒对鹿进行不同甲型疱疹病毒的筛查。我们比较了三种不同的商用ELISA试剂盒,以验证其在驯鹿和CvHV-2检测中的适用性。

方法

使用三种商用牛ELISA试剂盒(A、B和C),其中试剂盒A采用间接ELISA技术,试剂盒B和C采用阻断ELISA技术,检测针对BoHV-1的抗体,对154头驯鹿的血清进行检测,以检测针对CvHV-2的抗体。计算基于斯皮尔曼等级的相关系数(rho)。对所有试剂盒进行稀释试验。使用BoHV-1和CvHV-2进行病毒中和试验。

结果

所有试剂盒的血清阳性率几乎相同(40%-41%)。尽管定性评分相似,但在定量方面,试剂盒对样本的分类不同,仅在试剂盒B和C之间发现了强相关性。阻断试剂盒在重复性和稀释试验中表现更好。病毒中和结果与ELISA结果高度一致。针对CvHV-2和BoHV-1的中和效价分别为1:2至1:256和0至1:16。

结论

结果表明,BoHV-1和CvHV-2之间的基因和抗原相似性使得可以使用牛gB阻断ELISA试剂盒对驯鹿进行筛查。使用ELISA试剂盒既便宜又省时,能够对大量鹿群进行筛查。本研究发现大量动物对CvHV-2呈阳性反应,其在总体鹿群中的影响和分布应进一步评估。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e5f9/2663558/99b65dda510b/1751-0147-51-9-1.jpg

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