评价用于马匹驹年度分子监测的非侵入性采样技术。

Evaluation of Non-Invasive Sampling Techniques for the Molecular Surveillance of Equid Herpesviruses in Yearling Horses.

机构信息

Department of Veterinary Science, Martin-Gatton College of Agriculture, Food and the Environment, University of Kentucky, Lexington, KY 40506, USA.

Department of Public Health & Nutrition, University of Haripur, Haripur 22600, Pakistan.

出版信息

Viruses. 2024 Jul 7;16(7):1091. doi: 10.3390/v16071091.

DOI:10.3390/v16071091

PMID:39066254

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC11281437/

Abstract

BACKGROUND

Equid alphaherpesvirus 1 (EHV-1) is a highly contagious respiratory tract pathogen of horses, and infection may be followed by myeloencephalopathy or abortion. Surveillance and early detection have focused on PCR assays using less tolerated nasal swabs. Here, we assess non-invasive non-contact sampling techniques as surveillance tools in naturally equid gammaherpesvirus 2-shedding horses as surrogates for EHV-1.

METHODS

Horses were individually housed for 10 h periods on 2 consecutive days. Sampling included nasal swabs, nostril wipes, environmental swabs, droplet-catching devices, and air sampling. The latter was completed via two strategies: a combined air sample collected while going from horse to horse and a collective air sample collected at a stationary central point for 6 h. Samples were screened through quantitative PCR and digital PCR.

RESULTS

Nine horses on day 1 and 11 horses on day 2 were positive for EHV-1; overall, 90.9% of the nostril wipes, 81.8% of the environmental surfaces, and 90.9% of the droplet-catching devices were found to be positive. Quantitative analysis showed that the mean DNA copies detection per cm of nostril wipe sampled concentration (4.3 × 10 per day) was significantly ( < 0.05) comparable to that of nasal swabs (3.6 × 10 per day) followed by environmental swabs (4.3 × 10 per day) and droplet catchers (3.5 × 10 per day), respectively. Overall, 100% of the air samples collected were positive on both qPCR and dPCR. In individual air samples, a mean concentration of 1.0 × 10 copies of DNA were detected in per m air sampled per day, while in the collective air samples, the mean concentration was 1.1 × 10.

CONCLUSIONS

Environmental samples look promising in replacing direct contact sampling. Environmental and air sampling could become efficient surveillance tools at equestrian events; however, it needs threshold calculations for minimum detection levels.

摘要

背景

马疱疹病毒 1 型（EHV-1）是一种高度传染性的马呼吸道病原体，感染后可能导致脑脊髓炎或流产。监测和早期检测主要集中在使用耐受性较差的鼻腔拭子的 PCR 检测上。在这里，我们评估了非侵入性非接触式采样技术作为自然排放马疱疹病毒 2 型的马的监测工具，作为 EHV-1 的替代品。

方法

马在连续两天的 10 小时期间被单独关在一个房间里。采样包括鼻腔拭子、鼻孔擦拭物、环境拭子、液滴收集装置和空气采样。后者通过两种策略完成：一种是在从一匹马走到另一匹马时收集的综合空气样本，另一种是在一个固定的中心点收集 6 小时的集体空气样本。样本通过定量 PCR 和数字 PCR 进行筛选。

结果

第 1 天有 9 匹马和第 2 天有 11 匹马 EHV-1 阳性；总体而言，90.9%的鼻孔擦拭物、81.8%的环境表面和 90.9%的液滴收集器被发现呈阳性。定量分析表明，每厘米采样浓度的鼻腔拭子的平均 DNA 拷贝检测值（每天 4.3×10 拷贝）与鼻腔拭子（每天 3.6×10 拷贝）、环境拭子（每天 4.3×10 拷贝）和液滴收集器（每天 3.5×10 拷贝）相比，差异有统计学意义（<0.05）。总体而言，qPCR 和 dPCR 两种方法对空气样本的阳性检出率均为 100%。在个体空气样本中，每天每立方米空气中检测到的平均 DNA 拷贝数为 1.0×10 拷贝，而在集体空气样本中，平均浓度为 1.1×10。