用于检测犬血清中犬腺病毒2型抗体的间接酶联免疫吸附测定法的开发。

Development of indirect ELISA for the detection of canine adenovirus type 2 antibodies in dog sera.

作者信息

Yang Dong Kun, Kim Ha Hyun, Lee Siu, Oh Dongryul, Yoo Jae Young, Hyun Bang Hun

机构信息

Viral Disease Research Division, Animal and Plant Quarantine Agency (APQA), Ministry for Agriculture, Food and Rural Affairs (MAFRA), Gimcheon 39660, Korea.

出版信息

J Vet Sci. 2020 Jul;21(4):e63. doi: 10.4142/jvs.2020.21.e63.

DOI:10.4142/jvs.2020.21.e63

PMID:32735100

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC7402943/

Abstract

BACKGROUND

Canine adenovirus type 2 (CAV-2) induces infectious laryngotracheitis in members of the family Canidae, including dogs. To date, no ELISA kits specific for CAV-2 antibody have been commercialized for dogs in Korea.

OBJECTIVES

We aimed to develop new indirect enzyme-linked immunosorbent assay (I-ELISA) to perform rapid, accurate serological surveys of CAV-2 in dog serum samples.

METHODS

In total, 165 serum samples were collected from dogs residing in Chungbuk and Gyeongbuk provinces between 2016 and 2018. The Korean CAV-2, named the APQA1701-40P strain, was propagated in Madin-Darby canine kidney cells and purified in an anion-exchange chromatography column for use as an antigen for I-ELISA. The virus-neutralizing antibody titers of CAV-2 in the dog sera were measured by virus neutralization (VN) test.

RESULTS

We compared the results obtained between the VN and new I-ELISA tests. The sensitivity, specificity, and accuracy of new I-ELISA were 98.6%, 86.4% and 97.0% compared with VN test, respectively. New I-ELISA was significantly correlated with VN ( = 0.91).

CONCLUSIONS

These results indicate that new I-ELISA is useful for sero-surveillance of CAV-2 in dog serum.

摘要

背景

犬腺病毒2型（CAV - 2）可在包括犬科动物在内的犬科动物成员中引发传染性喉气管炎。迄今为止，韩国尚未有针对犬CAV - 2抗体的ELISA试剂盒商业化。

目的

我们旨在开发新的间接酶联免疫吸附测定法（I - ELISA），以便对犬血清样本中的CAV - 2进行快速、准确的血清学检测。

方法

2016年至2018年期间，共从忠清北道和庆尚北道的犬只中采集了165份血清样本。将命名为APQA1701 - 40P株的韩国CAV - 2在马迪 - 达比犬肾细胞中繁殖，并在阴离子交换色谱柱中纯化，用作I - ELISA的抗原。通过病毒中和（VN）试验测定犬血清中CAV - 2的病毒中和抗体效价。