Detection of Pathogens of Acute Febrile Illness Using Polymerase Chain Reaction from Dried Blood Spots.
Detection of Pathogens of Acute Febrile Illness Using Polymerase Chain Reaction from Dried Blood Spots.
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DOI：
10.4269/ajtmh.21-0814
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发表时间：
2021-11-08
期刊：
影响因子：
--
通讯作者：
Marks F
中科院分区：
文献类型：
--
作者：
Grundy B;Panzner U;Liu J;Jeon HJ;Im J;von Kalckreuth V;Konings F;Pak GD;Cruz Espinoza LM;Bassiahi AS;Gasmelseed N;Rakotozandrindrainy R;Stroup S;Houpt ER;Marks F
用于病原体检测的干血斑（DBS）定量聚合酶链反应（qPCR）是一种潜在的便捷的传染病诊断方法。这项研究通过 qPCR 测试了 115 个 DBS 样本以及来自布基纳法索、苏丹和马达加斯加的儿童和青少年的全血样本，检测了多种病原体，包括原生动物、蠕虫、真菌、细菌和病毒。疟原虫属从 DBS 中一致检测到，但产生的平均循环阈值 (Ct) 比全血样本高 5.7 ± 1.6。 DBS qPCR Ct 截止值 27 与之前建议用于疟疾诊断的全血 qPCR 截止值 21 相比，灵敏度为 94.1%，特异性为 95.1%。对于其他研究的病原体，与全血 qPCR 相比，DBS 检测的敏感性仅为 8.5%，但特异性为 98.6%。总之，DBS 的直接 PCR 对疟原虫具有合理的性能，但需要对本研究中评估的其他病原体进行进一步研究。
Quantitative polymerase chain reaction (qPCR) of dried blood spots (DBS) for pathogen detection is a potentially convenient method for infectious disease diagnosis. This study tested 115 DBS samples paired with whole blood specimens of children and adolescent from Burkina Faso, Sudan, and Madagascar by qPCR for a wide range of pathogens, including protozoans, helminths, fungi, bacteria, and viruses. Plasmodium spp. was consistently detected from DBS but yielded a mean cycle threshold (Ct) 5.7 ± 1.6 higher than that from whole blood samples. A DBS qPCR Ct cutoff of 27 yielded 94.1% sensitivity and 95.1% specificity against the whole blood qPCR cutoff of 21 that has been previously suggested for malaria diagnosis. For other pathogens investigated, DBS testing yielded a sensitivity of only 8.5% but a specificity of 98.6% compared with whole blood qPCR. In sum, direct PCR of DBS had reasonable performance for Plasmodium but requires further investigation for the other pathogens assessed in this study.
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