pfhrp2 and pfhrp3 Gene Deletions That Affect Malaria Rapid Diagnostic Tests for Plasmodium falciparum : Analysis of Archived Blood Samples From 3 African Countries

Abstract Background Malaria rapid diagnostic tests (mRDTs) that target histidine-rich protein 2 (HRP2) are important tools for Plasmodium falciparum diagnosis. Parasites with pfhrp2/3 gene deletions threaten the use of these mRDTs and have been reported in Africa, Asia, and South America. We studied blood samples from 3 African countries to determine if these gene deletions were present. Methods We analyzed 911 dried blood spots from Ghana (n = 165), Tanzania (n = 176), and Uganda (n = 570). Plasmodium falciparum infection was confirmed by 18S rDNA polymerase chain reaction (PCR), and pfhrp2/3 genes were genotyped. True pfhrp2/3 gene deletions were confirmed if samples were (1) microscopy positive; (2) 18S rDNA PCR positive; (3) positive for merozoite surface protein genes by PCR or positive by loop-mediated isothermal amplification; or (4) quantitative PCR positive with >5 parasites/µL. Results No pfhrp2/3 deletions were detected in samples from Ghana, but deletions were identified in Tanzania (3 pfhrp2 ; 2 pfhrp3 ) and Uganda (7 pfhrp2 ; 2 pfhrp3 ). Of the 10 samples with pfhrp2 deletions, 9 tested negative by HRP2-based mRDT. Conclusions The presence of pfhrp2/3 deletions in Tanzania and Uganda, along with reports of pfhrp2/3 -deleted parasites in neighboring countries, reinforces the need for systematic surveillance to monitor the reliability of mRDTs in malaria-endemic countries. pfhrp2 gene deletions render Plasmodium falciparum parasites undetectable to malaria rapid diagnostic tests detecting histidine-rich protein 2. pfhrp2 deletions were detected in archived blood samples from Tanzania and Uganda, while no samples from Ghana were found to be pfhpr2 -negative.