A novel method for extracting nucleic acids from dried blood spots for ultrasensitive detection of low-density Plasmodium falciparum and Plasmodium vivax infections.
A novel method for extracting nucleic acids from dried blood spots for ultrasensitive detection of low-density Plasmodium falciparum and Plasmodium vivax infections.
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DOI：
10.1186/s12936-017-2025-3
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发表时间：
2017-09-18
期刊：
影响因子：
3
通讯作者：
Nyunt MM
中科院分区：
文献类型：
--
作者：
Zainabadi K;Adams M;Han ZY;Lwin HW;Han KT;Ouattara A;Thura S;Plowe CV;Nyunt MM
大湄公河次区域各国承诺到2025年消除恶性疟原虫疟疾。目前的消除干预措施针对的是可以通过标准显微镜或快速诊断测试检测到的寄生虫密度的感染。已经开发了更灵敏的检测方法，以检测可能代表重要传播宿主的较低密度“无症状”感染。这些超灵敏的聚合酶链反应（usPCR）测试已被用于确定大规模药物管理（MDA）的目标人群。迄今为止，疟疾usPCR检测使用静脉或毛细血管血液采样，这需要复杂的样本收集、处理和运输要求。对标准干血斑（DBS）进行的超灵敏方法将极大地促进靶向消除干预所需的分子监测研究。通过优化核酸提取条件，建立了一种高灵敏度的检测DBS中恶性疟原虫和间日疟原虫18 S核糖体RNA的方法。使用在模拟现场条件下干燥和储存的加标DBS样品测定检测限（LoD）。此外，为了评估其在常规分子监测中的效用，在缅甸的雨季和旱季进行了两次横断面调查。基于DBS的超灵敏检测试剂的LoD下限为20个寄生虫/mL（对于Whatman 3 MM滤纸上采集的DBS）和23个寄生虫/mL（对于Whatman 903 Protein Saver卡）-相当于每50 µL DBS 1个寄生虫。这比标准RDT的灵敏度高约5000倍，与基于全血的其他超灵敏方法报告的LoD（≤16-22个寄生虫/mL）相似。在缅甸的两项横断面调查中，从同期DBS样本和在雨季和旱季采集的毛细血管血液样本中获得了几乎相同的患病率估计值。本研究中描述的基于DBS的超灵敏方法在两次实地调查中的检测限和患病率估计值方面与先前描述的基于全血的方法具有相同的灵敏度。这种方法降低了成本和复杂性，将能够扩大监测研究的规模，以针对MDA和其他消除疟疾的干预措施，并有助于更好地了解低密度疟疾感染的流行病学。本文的在线版本（doi：10.1186/s12936-017-2025-3）包含补充材料，可供授权用户使用。
Greater Mekong Subregion countries are committed to eliminating Plasmodium falciparum malaria by 2025. Current elimination interventions target infections at parasite densities that can be detected by standard microscopy or rapid diagnostic tests (RDTs). More sensitive detection methods have been developed to detect lower density “asymptomatic” infections that may represent an important transmission reservoir. These ultrasensitive polymerase chain reaction (usPCR) tests have been used to identify target populations for mass drug administration (MDA). To date, malaria usPCR tests have used either venous or capillary blood sampling, which entails complex sample collection, processing and shipping requirements. An ultrasensitive method performed on standard dried blood spots (DBS) would greatly facilitate the molecular surveillance studies needed for targeting elimination interventions. A highly sensitive method for detecting Plasmodium falciparum and P. vivax 18S ribosomal RNA from DBS was developed by empirically optimizing nucleic acid extraction conditions. The limit of detection (LoD) was determined using spiked DBS samples that were dried and stored under simulated field conditions. Further, to assess its utility for routine molecular surveillance, two cross-sectional surveys were performed in Myanmar during the wet and dry seasons. The lower LoD of the DBS-based ultrasensitive assay was 20 parasites/mL for DBS collected on Whatman 3MM filter paper and 23 parasites/mL for Whatman 903 Protein Saver cards—equivalent to 1 parasite per 50 µL DBS. This is about 5000-fold more sensitive than standard RDTs and similar to the LoD of ≤16–22 parasites/mL reported for other ultrasensitive methods based on whole blood. In two cross-sectional surveys in Myanmar, nearly identical prevalence estimates were obtained from contemporaneous DBS samples and capillary blood samples collected during the wet and dry season. The DBS-based ultrasensitive method described in this study shows equal sensitivity as previously described methods based on whole blood, both in its limit of detection and prevalence estimates in two field surveys. The reduced cost and complexity of this method will allow for the scale-up of surveillance studies to target MDA and other malaria elimination interventions, and help lead to a better understanding of the epidemiology of low-density malaria infections. The online version of this article (doi:10.1186/s12936-017-2025-3) contains supplementary material, which is available to authorized users.
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影响因子：
15.8
作者：
Chen I;Clarke SE;Gosling R;Hamainza B;Killeen G;Magill A;O'Meara W;Price RN;Riley EM
通讯作者：
Riley EM
柬埔寨恶性疟原虫疟疾中的双氢青蒿素-哌喹抗药性：一项多地点前瞻性队列研究。
DOI：
10.1016/s1473-3099(15)00487-9
发表时间：
2016-03
期刊：
The Lancet. Infectious diseases
影响因子：
--
作者：
Amaratunga C;Lim P;Suon S;Sreng S;Mao S;Sopha C;Sam B;Dek D;Try V;Amato R;Blessborn D;Song L;Tullo GS;Fay MP;Anderson JM;Tarning J;Fairhurst RM
通讯作者：
Fairhurst RM
影响因子：
3.7
作者：
Kemleu S;Guelig D;Eboumbou Moukoko C;Essangui E;Diesburg S;Mouliom A;Melingui B;Manga J;Donkeu C;Epote A;Texier G;LaBarre P;Burton R;Ayong L
通讯作者：
Ayong L
影响因子：
9.4
作者：
Kamau, Edwin;Tolbert, LaDonna S.;Ockenhouse, Christian F.
通讯作者：
Ockenhouse, Christian F.
影响因子：
9.4
作者：
Murphy, Sean C.;Daza, Glenda;Coombs, Robert
通讯作者：
Coombs, Robert