Rapid Detection of Actinobacillus pleuropneumoniae From Clinical Samples Using Recombinase Polymerase Amplification.
Rapid Detection of Actinobacillus pleuropneumoniae From Clinical Samples Using Recombinase Polymerase Amplification.
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DOI：
10.3389/fvets.2022.805382
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发表时间：
2022
影响因子：
3.2
通讯作者：
Langford PR
中科院分区：
文献类型：
--
作者：
Stringer OW;Li Y;Bossé JT;Forrest MS;Hernandez-Garcia J;Tucker AW;Nunes T;Costa F;Mortensen P;Velazquez E;Penny P;Rodriguez-Manzano J;Georgiou P;Langford PR
胸膜肺炎放线杆菌（Actinobacilluspleuropneumoniae，APP）是引起猪胸膜肺炎的病原菌，在世界范围内造成了巨大的经济损失。目前已知的APP血清型有19种，不同的血清型在特定的地理区域占主导地位。胸膜肺炎的爆发，其特征是突然的呼吸困难和高死亡率，当受感染的猪被带入未感染的猪群时，或被携带不同血清型的猪群感染时，可能会发生。良好的生物安保措施包括为监测目的定期进行诊断检测。目前的黄金标准诊断技术缺乏灵敏度（细菌培养），需要昂贵的热循环机械（PCR），并且耗时（培养和PCR）。在这里，我们描述了一种等温床旁诊断测试的发展-利用重组酶聚合酶扩增（RPA）检测APP，针对物种特异性apxIVA基因。我们的APP-RPA诊断测试使用APP血清型8菌株实现了10拷贝/μL的灵敏度，该菌株是英国最流行的血清型。此外，我们的APP-RPA检测在从英格兰和葡萄牙农场获得的61份提取的临床样本中分别实现了84.3%和100%的临床灵敏度和特异性。使用肺组织样本的一个小子集（n = 14），我们使用在APP-RPA反应中直接测试的FTA卡上制备的肺印记分别实现了76.9%和100%的临床灵敏度和特异性。我们的研究结果表明，我们的APP-RPA检测能够为这种重要的兽医病原体提供合适的快速和灵敏的筛查工具。
Actinobacillus pleuropneumoniae (APP) is the causative agent of porcine pleuropneumonia, resulting in high economic impact worldwide. There are currently 19 known serovars of APP, with different ones being predominant in specific geographic regions. Outbreaks of pleuropneumonia, characterized by sudden respiratory difficulties and high mortality, can occur when infected pigs are brought into naïve herds, or by those carrying different serovars. Good biosecurity measures include regular diagnostic testing for surveillance purposes. Current gold standard diagnostic techniques lack sensitivity (bacterial culture), require expensive thermocycling machinery (PCR) and are time consuming (culture and PCR). Here we describe the development of an isothermal point-of-care diagnostic test - utilizing recombinase polymerase amplification (RPA) for the detection of APP, targeting the species-specific apxIVA gene. Our APP-RPA diagnostic test achieved a sensitivity of 10 copies/μL using a strain of APP serovar 8, which is the most prevalent serovar in the UK. Additionally, our APP-RPA assay achieved a clinical sensitivity and specificity of 84.3 and 100%, respectively, across 61 extracted clinical samples obtained from farms located in England and Portugal. Using a small subset (n = 14) of the lung tissue samples, we achieved a clinical sensitivity and specificity of 76.9 and 100%, respectively) using lung imprints made on FTA cards tested directly in the APP-RPA reaction. Our results demonstrate that our APP-RPA assay enables a suitable rapid and sensitive screening tool for this important veterinary pathogen.
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影响因子：
2.5
作者：
Opriessnig, T.;Gimenez-Lirola, L. G.;Halbur, P. G.
通讯作者：
Halbur, P. G.
影响因子：
9.4
作者：
MITTAL, KR
通讯作者：
MITTAL, KR
DOI：
10.12968/live.2020.25.6.308
发表时间：
2020-01-01
期刊：
Livestock
影响因子：
--
作者：
Gale, Christina;Velazquez, Eduardo
通讯作者：
Velazquez, Eduardo
DOI：
10.1590/s0100-736x2012000800014
发表时间：
2012-08-01
期刊：
Pesquisa Veterinária Brasileira
影响因子：
--
作者：
Ji, Hongwei;Li, Haitao;Xu, Zhiwen
通讯作者：
Xu, Zhiwen
影响因子：
5.5
作者：
O'Neilla C;Jones SCP;Bossé JT;Watson CM;Williamson SM;Rycroft AN;Simon Kroll J;Hartley HM;Langford PR
通讯作者：
Langford PR