Targeting multicopy prophage genes for the differential diagnosis of Lyme disease
Targeting multicopy prophage genes for the differential diagnosis of Lyme disease
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DOI：
10.1101/2020.12.02.20241687
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发表时间：
2020-12
期刊：
影响因子：
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通讯作者：
J. Shan;Ying Jia;L. Teulières;F. Patel;M. Clokie
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文献类型：
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作者：
J. Shan;Ying Jia;L. Teulières;F. Patel;M. Clokie
莱姆病（LD）的成功治疗需要准确的诊断测试;然而，大多数测试是不敏感和非特异性的。为了克服这些挑战，我们开发并验证了一种内部控制的定量PCR（Ter-qPCR），其靶向由仅在LD致病细菌中发现的原噬菌体编码的多拷贝末端酶大亚基（terL）基因。terL蛋白质帮助它们包装DNA。引人注目的是，Ter-qPCR的检测限经分析估计为22个拷贝和细菌掺入血液中的一个细菌细胞。此外，在健康个体和患有早期或晚期疾病的患者中检测到的terL的量方面存在显著的定量差异。总之，这些数据表明，靶向原噬菌体的PCR具有显著的能力来提供LD的鉴别诊断。原噬菌体编码的标记物在许多其他病原菌中普遍存在，使得这种方法高度适用于一般的细菌鉴定，可能彻底改变疾病的检测。
The successful treatment of Lyme disease (LD) requires an accurate diagnostic test; however, most tests are insensitive and unspecific. To overcome these challenges, we developed and validated an internally-controlled quantitative PCR (Ter-qPCR) that targets the multicopy terminase large subunit (terL) gene encoded by prophages that are only found in LD-causing bacteria. The terL protein helps phages pack their DNA. Strikingly, the detection limit of the Ter-qPCR was analytically estimated to be 22 copies and one bacterial cell in bacteria spiked blood. Furthermore, significant quantitative differences in terms of the amount of terL detected in healthy individuals and patients with either early or late disease. Together, the data suggests that the prophage-targeting PCR has significant power to provide a differential diagnosis for LD. Prophage encoded markers are prevalent in many other pathogenic bacteria rendering this approach highly applicable to bacterial identification in general, potentially revolutionising the detection of disease.