Feasibility of community at-home dried blood spot collection combined with pooled reverse transcription PCR as a viable and convenient method for malaria epidemiology studies.
Feasibility of community at-home dried blood spot collection combined with pooled reverse transcription PCR as a viable and convenient method for malaria epidemiology studies.
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社区在家干血点收集的可行性与汇总逆转录PCR相结合，作为疟疾流行病学研究的可行且方便的方法。
DOI：
10.1186/s12936-022-04239-x
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发表时间：
2022-07-14
期刊：
影响因子：
3
通讯作者：
Murphy, Sean C.
中科院分区：
文献类型：
--
作者：
Hergott, Dianna E. B.;Owalla, Tonny J.;Balkus, Jennifer E.;Apio, Bernadette;Lema, Jimmy;Cemeri, Barbara;Akileng, Andrew;Seilie, Annette M.;Chavtur, Chris;Staubus, Weston;Chang, Ming;Egwang, Thomas G.;Murphy, Sean C.
流行地区的许多疟原虫感染的密度低于标准诊断工具的检测限。这些感染威胁到控制工作，并可能影响疫苗和治疗药物的研究。需要简单，成本效益的方法来研究无症状的亚显微寄生虫病的自然史。使用合并和个体定量逆转录聚合酶链反应（qRT-PCR）分析的自我收集的干血斑（DBS）提供了这样的解决方案。在此，研究了每日在家DBS采集用于qRT-PCR的可行性和可接受性，以更好地了解低密度感染。2021年4月至5月期间，在乌干达东北部Katakwi区招募了快速诊断测试（RDT）阴性个体。在入组时和每周4次门诊访视时采集静脉血样和临床采集的DBS。参与者接受DBS采集培训，并要求在诊所访视之间每周采集6次DBS。在最终研究访视时，使用每日日记卡和Likert量表调查征求关于收集过程的意见。通过疟原虫18 S rRNA qRT-PCR分析静脉血和DBS。分析完成研究的参与者数量、收集的DBS总量以及对过程的意见，以确定依从性和可接受性。评价了在家与诊所采集的DBS和静脉血的人内部对照mRNA和疟原虫18 S rRNA，以评估在家采集样本的质量和准确度。102名成人和29名儿童入组，分别有95名和26名完成了研究。3人因疼痛或手术不便退出。总体而言，96%的参与者采集了24个家庭DBS中的≥ 16个，87%的DBS含有≥ 40 µL的血液。这一程序得到了与会者的良好容忍和好评。在家中收集的DBS对于qRT-PCR是可接受的，并且与临床收集的DBS相比，在人对照mRNA中显示出小于一个qRT-PCR循环的阈值偏移。配对全血中的恶性疟原虫18 S rRNA与DBS之间的相关性很高（R = 0.93）。家庭DBS采集是一种可行、可接受和稳健的方法，可用于采集血液，通过qRT-PCR评价低密度疟原虫感染的自然史。在线版本包含补充材料，可通过10.1186/s12936-022-04239-x获得。
Many Plasmodium infections in endemic regions exist at densities below the limit of detection of standard diagnostic tools. These infections threaten control efforts and may impact vaccine and therapeutic drug studies. Simple, cost-effective methods are needed to study the natural history of asymptomatic submicroscopic parasitaemia. Self-collected dried blood spots (DBS) analysed using pooled and individual quantitative reverse transcription polymerase chain reaction (qRT-PCR) provide such a solution. Here, the feasibility and acceptability of daily at-home DBS collections for qRT-PCR was studied to better understand low-density infections. Rapid diagnostic test (RDT)-negative individuals in Katakwi District, northeastern Uganda, were recruited between April and May 2021. Venous blood samples and clinic-collected DBS were taken at enrollment and at four weekly clinic visits. Participants were trained in DBS collection and asked to collect six DBS weekly between clinic visits. Opinions about the collection process were solicited using daily Diary Cards and a Likert scale survey at the final study visit. Venous blood and DBS were analysed by Plasmodium 18S rRNA qRT-PCR. The number of participants completing the study, total DBS collected, and opinions of the process were analysed to determine compliance and acceptability. The human internal control mRNA and Plasmodium 18S rRNA were evaluated for at-home vs. clinic-collected DBS and venous blood to assess quality and accuracy of at-home collected samples. One-hundred two adults and 29 children were enrolled, and 95 and 26 completed the study, respectively. Three individuals withdrew due to pain or inconvenience of procedures. Overall, 96% of participants collected ≥ 16 of 24 at-home DBS, and 87% of DBS contained ≥ 40 µL of blood. The procedure was well tolerated and viewed favourably by participants. At-home collected DBS were acceptable for qRT-PCR and showed less than a one qRT-PCR cycle threshold shift in the human control mRNA compared to clinic-collected DBS. Correlation between Plasmodium falciparum 18S rRNA from paired whole blood and DBS was high (R = 0.93). At-home DBS collection is a feasible, acceptable, and robust method to obtain blood to evaluate the natural history of low-density Plasmodium infections by qRT-PCR. The online version contains supplementary material available at 10.1186/s12936-022-04239-x.
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影响因子：
3
作者：
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通讯作者：
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影响因子：
3.3
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