Reverse transcription PCR to detect low density malaria infections

逆转录 PCR 检测低密度疟疾感染

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作者:Peter Christensen, Zbynek Bozdech, Wanitda Watthanaworawit, Laurent Rénia, Benoît Malleret, Clare Ling, François Nosten

Background

Targeted malaria elimination strategies require highly sensitive tests to detect low density malaria infections (LDMI). Commonly used

Conclusion

Malaria detection in areas of low transmission and LDMI can benefit from the increased sensitivity of ribosomal RNA detection by RT-PCR, especially where sample volume is limited. Isolation of high quality RNA also allows for downstream analysis of malaria transcripts.

Methods

Here we evaluated a real-time reverse transcription PCR (RT-qPCR) method that 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 RT-qPCR.

Results

Plasmodium spp. RT-qPCR confirmed 18 of 21 uPCR Plasmodium falciparum positives, while P. falciparum specific RT-qPCR confirmed 17 of the 21 uPCR P. falciparum positives. Combining both RT-qPCR results increased the sensitivity to 100% and specificity was 95.1%.

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