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提高非洲马病毒检测:比较和调整PCR检测方法

Lisa Penzhorn¹, Jan E Crafford², Alan J Guthrie¹

¹Equine Research Centre, Faculty of Veterinary Science, University of Pretoria, Onderstepoort, South Africa.

Journal of veterinary diagnostic investigation : official publication of the American Association of Veterinary Laboratory Diagnosticians, Inc

|February 7, 2026

概括

通过修改RT-qPCR试验,提高了非洲马病病毒 (AHSV) 检测准确度. 更新的测试实现了100%的灵敏度,增强了对这种严重的马匹威胁的诊断和疾病控制.

科学领域:

兽医病毒学兽医病毒学
分子诊断学分子诊断
疾病监测疾病监测

背景情况:

非洲马病 (AHS) 是对马业的重大威胁,该病毒 (AHSV) 流行于撒哈拉以南非洲.
精确诊断AHSV对于疾病控制和国际贸易至关重要,RT-qPCR是关键的诊断工具.
在AHSV的遗传变异可能会影响当前RT-qPCR测定的准确性,可能导致假阴性.

研究的目的:

评估WOAH推的用于检测AHSV的RT-qPCR试验的诊断准确性.
为了识别现有测试中由于AHSV遗传变异性而导致的潜在失败.
开发和验证改进的RT-qPCR测定,以提高AHSV检测的灵敏度和准确性.

主要方法:

在实验室分析150个AHSV阳性样本,使用Agüero等. 和古特里等人. 进行RT-qPCR测定.
从假阴性样本和公开存档的序列中对AHSV VP7基因进行in silico分析.
根据in silico发现,在Guthrie和Agüero试验中对原料和探针的修改.
第二次体外分析,以评估修改后的测试的诊断准确性.

主要成果:

原始的Agüero检测显示出8.7%的假阴性率,未能在13个样本中检测到AHSV.
在基分析中发现了影响原料/探针结合的遗传变异.
经过修改的Agüero和Guthrie测定在第二次体外分析中显示出100%的灵敏度.
在体和体外的结果都强调了需要持续监测RT-qPCR疗效的必要性.

结论:

现有的WOAH推的AHSV的RT-qPCR测定可能由于病毒遗传变异性而导致诊断准确性的局限性.
对原料和探针的修改可以显著提高RT-qPCR测定用于AHSV检测的灵敏度.
持续监测和更新分子诊断协议对于准确的AHSV监测和控制至关重要.

关键词:

非洲马病是非洲马病.反向转录实时PCR进行反转录.测试的准确性测试的准确性

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