Targeted malaria elimination strategies require highly sensitive tests to detect low density malaria infections (LDMI). Commonly used methods for malaria diagnosis such as light microscopy and antigen-based rapid diagnostic tests (RDTs) are not sensitive enough for reliable identification of infections with parasitaemia below 200 parasites per milliliter of blood. While targeted malaria elimination efforts on the Thailand-Myanmar border have successfully used high sample volume ultrasensitive quantitative PCR (uPCR) to determine malaria prevalence, the necessity for venous collection and processing of large quantities of patient blood limits the widespread tractability of this method. Here we evaluated a real-time quantitative reverse transcription PCR (qRT-PCR) method that significantly reduces the required sample volume compared to uPCR. To do this, 304 samples collected from an active case detection program in Kayin state, Myanmar were compared using uPCR and qRT-PCR. Plasmodium spp. qRT-PCR confirmed 18 of 21 uPCR Plasmodium falciparum positives, while P. falciparum specific qRT-PCR confirmed 17 of the 21 uPCR P. falciparum positives. Combining both qRT-PCR results increased the sensitivity to 100% and specificity was 95.1%. These results show that malaria detection in areas of low transmission and LDMI, can benefit from the increased sensitivity of qRT-PCR especially where sample volume is limited.

中文翻译：

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逆转录定量PCR检测低密度疟疾感染

有针对性的消除疟疾策略需要高度敏感的测试，以检测低密度疟疾感染（LDMI）。常用的疟疾诊断方法（如光学显微镜和基于抗原的快速诊断检测（RDT））不够敏感，无法可靠地鉴定每毫升血液中寄生虫病低于200寄生虫的感染。尽管在泰国-缅甸边境开展的针对性疟疾消除工作已成功地使用高样本量超灵敏定量PCR（uPCR）来确定疟疾患病率，但静脉收集和处理大量患者血液的必要性限制了该方法的广泛可扩展性。在这里，我们评估了实时定量逆转录PCR（qRT-PCR）方法，与uPCR相比，该方法大大减少了所需的样品量。为此，使用uPCR和qRT-PCR对从缅甸卡音州一个活跃病例检测程序收集的304个样本进行了比较。疟原虫属 qRT-PCR确认了21个uPCR恶性疟原虫阳性中的18个，而恶性疟原虫特异性qRT-PCR确认了21个uPCR恶性疟原虫阳性中的17个。结合两个qRT-PCR结果，将灵敏度提高到100％，特异性为95.1％。这些结果表明，在低透射率和LDMI区域中进行疟疾检测可以受益于qRT-PCR灵敏度的提高，尤其是在样品量有限的情况下。恶性疟原虫。结合两个qRT-PCR结果，将灵敏度提高到100％，特异性为95.1％。这些结果表明，在低透射率和LDMI区域中进行疟疾检测可以受益于qRT-PCR灵敏度的提高，尤其是在样品量有限的地方。恶性疟原虫。结合两个qRT-PCR结果，将灵敏度提高到100％，特异性为95.1％。这些结果表明，在低透射率和LDMI区域检测疟疾可以受益于qRT-PCR灵敏度的提高，尤其是在样品量有限的地区。