Molecular surveillance of pfhrp2 and pfhrp3 genes deletion in Plasmodium falciparum isolates and the implications for rapid diagnostic tests in Nigeria

Funwei, Roland I.; Nderu, David; Nguetse, Christian N.; Thomas, Bolaji N.; Falade, Catherine O.; Velavan, Thirumalaisamy P.; Ojurongbe, Olusola

doi:10.1016/j.actatropica.2019.05.016

Cited by 46 publications

(48 citation statements)

References 42 publications

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“…Only 9.9% of false-negative PfHRP2-RDT results involved parasites with pfhrp2 gene deletions. A similar result (10.6%) has been reported in Nigeria, (24). Conversely, Wurtz et al (2013), in Senegal reported a lower proportion (2.4%) while Koita et al (2012), in Mali and Amoah et al (2016), in Ghana reported higher proportions, 45% and 23%, respectively (4,5,17).…”

Section: Discussionsupporting

confidence: 72%

Prevalence of Plasmodium falciparum Isolates Lacking the Histidine Rich Protein 2 Gene Among Symptomatic Malaria Patients in Kwilu Province, DR. Congo.

Munyeku¹,

Musaka²,

Ernest³

et al. 2020

Preprint

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BackgroundMalaria rapid diagnostic tests have become a primary and critical tool for malaria diagnosis in malaria-endemic countries where PfHRP2-based RDTs are widely used. However, in the last decade, the accuracy of PfHRP2-based RDTs has been challenged by the emergence of P. falciparum strains harbouring deletions of the pfhrp2 gene, resulting in false-negative results. In DR. Congo, little is known about the prevalence of the pfhrp2 gene deletion among P. falciparum isolates infecting symptomatic patients, especially in low to moderate transmission areas where pfhrp2 deletion parasites are assumed to emerge and spread. Here we determine the local prevalence and factors associated with pfhrp2 gene deletions among symptomatic malaria patients in the Kwilu province of the Democratic Republic of Congo, a low to moderate malaria transmission area.MethodsWe used secondary data from a prospective health facility-based cross-sectional study conducted on 684 individuals of all ages, seeking healthcare with symptoms suggestive of malaria from October to December 2018 in 34 randomly selected health facilities. Sociodemographic, malaria prevention and treatment practices, and clinical variables were collected using a pre-tested structured questionnaire. Patients’ medical records were used to collect additional clinical data. Blood was collected for microscopy, PfHRP2-RDT, and spotted onto Whatman filter paper for downstream genetic analysis. Genomic DNA was extracted and used to perform PCR assays for the detection and confirmation of pfhrp2 gene deletions. Data were entered and analysed using STATA15. Fischer’s exact and the Kruskal-Wallis tests were applied to look for associations between exposures and the pfhrp2 gene deletion with a level of statistical significance set at p < 0.05. ResultsThe overall prevalence of the pfhrp2 gene deletion was 9.2% (95% CI 6.7% – 12.1%). The deletion of the pfhrp2 gene was associated with health zone of origin (p=0.012) and age (p=0.019). Among false-negative PfHRP2-RDT results, only 9.9% were due to pfhrp2 gene deletion.ConclusionP. falciparum isolates with pfhrp2 gene deletions are relatively common among symptomatic patients in Kwilu province. Further investigations are needed to provide enough evidence for policy change. Meanwhile, the use of RDTs targeting PfHRP2 and pLDH antigens could limit the spread of deleted isolates.

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Section: Discussionsupporting

confidence: 72%

Prevalence of Plasmodium falciparum Isolates Lacking the Histidine Rich Protein 2 Gene Among Symptomatic Malaria Patients in Kwilu Province, DR. Congo.

Munyeku¹,

Musaka²,

Ernest³

et al. 2020

Preprint

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“…Similarly, the majority of Myanmar pfhrp3 started with type 1 repeat and terminated with type 4 repeat. This conserved structural organization was also identified in global pfhrp2 and pfhrp3 [ 25 , 27 – 29 ]; the major repeat types found in Myanmar pfhrp2 and pfhrp3 were also the most common repeat types observed in global pfhrp2 and pfhrp3 . Although the frequency of each repeat type differed slightly in global isolates, repeat types 1, 2, 3, 6, 7, 8, 10, and 12 were the most common in Myanmar and global pfhrp2 [ 25 , 27 – 29 ].…”

Section: Discussionmentioning

confidence: 58%

Genetic variations in histidine-rich protein 2 and histidine-rich protein 3 of Myanmar Plasmodium falciparum isolates

Lê

Kang

Lee

et al. 2020

Malar J

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Background Malaria rapid diagnostic tests (RDTs) are precious tools to diagnose malaria. Most RDTs used currently are based on the detection of Plasmodium falciparum histidine-rich protein 2 (PfHRP2) in a patient’s blood. However, concern has been raised in recent years that deletion of pfhrp2 in the parasite could affect the accuracy of PfHRP2-based RDTs. In addition, genetic variation in pfhrp2 might influence the accuracy and sensitivity of RDTs. In this study, the genetic variation in pfhrp2 and pfhrp3 in Myanmar P. falciparum isolates was analysed. Methods Blood samples were collected from malaria patients who were infected with P. falciparum in Mandalay, Naung Cho, Tha Beik Kyin, and Pyin Oo Lwin, Upper Myanmar between 2013 and 2015. The pfhrp2 and pfhrp3 were amplified by nested polymerase chain reaction (PCR), cloned and sequenced. Genetic variation in Myanmar pfhrp2 and pfhrp3 was analysed using the DNASTAR program. Comparative analysis of Myanmar and global pfhrp2 and pfhrp3 isolates was also performed. Results One-hundred and two pfhrp2 and 89 pfhrp3 were amplified from 105 blood samples, of which 84 pfhrp2 and 56 pfhrp3 sequences were obtained successfully. Myanmar pfhrp2 and pfhrp3 showed high levels of genetic variation with different arrangements of distinct repeat types, which further classified Myanmar pfhrp2 and pfhrp3 into 76 and 47 haplotypes, respectively. Novel amino acid changes were also found in Myanmar pfhrp2 and pfhrp3, but their frequencies were very low. Similar structural organization was shared by Myanmar and global pfhrp2 and pfhrp3, and differences in frequencies of repeat types and lengths were also observed between and among global isolates. Conclusion Length polymorphisms and amino acid substitutions generated extensive genetic variation in Myanmar pfhrp2 and pfhrp3. Comparative analysis revealed that global pfhrp2 and pfhrp3 share similar structural features, as well as extensive length polymorphisms and distinct organizations of repeat types. These results provide a better understanding of the genetic structure of pfhrp2 and pfhrp3 in global P. falciparum populations and suggest useful information to develop RDTs with improved quality.

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“…Eight studies were excluded from the analysis on the proportion of P. falciparum isolates with deletions in pfhrp2/3 genes among false negative PfHRP2-based RDT results because of small sample size and the topic not being addressed. Sixteen and 10 studies were included in the meta-analysis to determine the prevalence of deletions in pfhrp2/3 genes and their proportion among false negative PfHRP2-based RDT results, respectively [26][27][28][29][30][31][32][33][34][35][36][37][38][39][40][41][42][43][44] (Fig. 1, Additional file 2).…”

Section: Selection Of Studies Included In the Meta-analysismentioning

confidence: 99%

“…The 16 studies from 13 SSA countries: Mali, Senegal, Ghana, Kenya, Mozambique, Rwanda, Zambia, Eritrea, Eswatini (Swaziland), Nigeria, Sudan, Madagascar and Democratic Republic of Congo, were conducted locally with one exception which was conducted at national level ( Table 1) [26][27][28][29][30][31][32][33][34][35][36][37][38][39][40][41][42][43][44]. Three studies found deletions in pfhrp2 in Uganda, Gambia and Tanzania [45][46][47], but these were excluded from the systematic review (Additional file 5).…”

Section: Characteristics Of Studies Included In the Meta-analysismentioning

confidence: 99%

Prevalence of Plasmodium falciparum field isolates with deletions in histidine-rich protein 2 and 3 genes in context with sub-Saharan Africa and India: a systematic review and meta-analysis

Kojom

Singh

2020

Malar J

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Background: In 2017, nearly 80% of malaria morbidity and mortality occurred in sub-Saharan African (SSA) countries and India. Rapid diagnostic tests (RDTs), especially those targeting histidine-rich protein 2 (PfHRP2) of Plasmodium falciparum, have become an important diagnostic tool in these malaria-endemic areas. However, the chances of RDToriented successful treatment are increasingly jeopardized by the appearance of mutants with deletions in pfhrp2 and pfhrp3 genes. This systematic review and meta-analysis determines the prevalence of field P. falciparum isolates with deletion in pfhrp2 and/or pfhrp3 genes and their proportion among false-negative results in the PfHRP2-based RDTs in SSA and India.Methods: Eight electronic databases were used for searching potentially relevant publications for the systematic review analysis, wherein the main methodological aspects of included studies were analysed and some missing links in the included studies were identified. Results:A total of 19 studies were included, 16 from SSA and 3 from India. The pooled prevalence of pfhrp2 deletions was 8 and 5% while 16 and 4% for pfhrp3 gene deletions in Africa and India, respectively. The pooled proportion of pfhrp2 gene deletions found among false negative PfHRP2-based RDTs results was about 27.0 and 69.0% in Africa and India, respectively. Conclusions:This review study indicates a relatively high proportion of both pfhrp2/3 genes deletions in P. falciparum isolates and among false-negative malaria cases using PfHRP2-based RDT results in SSA and India. Recently the deletions in pfhrp2/3 genes have also been reported from two African countries (Nigeria and Sudan). This review emphasizes the importance of more extensive studies and standardization of studies addressing the pfhrp2/3 gene deletions in malarious areas.

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Molecular surveillance of pfhrp2 and pfhrp3 genes deletion in Plasmodium falciparum isolates and the implications for rapid diagnostic tests in Nigeria

Cited by 46 publications

References 42 publications

Prevalence of Plasmodium falciparum Isolates Lacking the Histidine Rich Protein 2 Gene Among Symptomatic Malaria Patients in Kwilu Province, DR. Congo.

Prevalence of Plasmodium falciparum Isolates Lacking the Histidine Rich Protein 2 Gene Among Symptomatic Malaria Patients in Kwilu Province, DR. Congo.

Genetic variations in histidine-rich protein 2 and histidine-rich protein 3 of Myanmar Plasmodium falciparum isolates

Prevalence of Plasmodium falciparum field isolates with deletions in histidine-rich protein 2 and 3 genes in context with sub-Saharan Africa and India: a systematic review and meta-analysis

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