Molecular identification and anti-malarial drug resistance profile of Plasmodium falciparum from patients attending Kisoro Hospital, southwestern Uganda

Abstract

Background The evolution of malaria infection has necessitated the development of highly sensitive diagnostic assays, as well as the use of dried blood spots (DBS) as a potential source of deoxyribonucleic acid (DNA) yield for polymerase chain reaction (PCR) assays. This study identified the different Plasmodium species in malaria-positive patients, and the anti-malarial drug resistance profile for Plasmodium falciparum using DBS samples collected from patients attending Kisoro Hospital in Kisoro district, Southwestern Uganda. Methods The blood samples were prospectively collected from patients diagnosed with malaria to make DBS, which were then used to extract DNA for real-time PCR and high-resolution melting (HRM) analysis. Plasmodium species were identified by comparing the control and test samples using HRM-PCR derivative curves. Plasmodium falciparum chloroquine (CQ) resistance transporter (pfcrt) and kelch13 to screen the samples for anti-malarial resistance markers. The HRM-PCR derivative curve was used to present a summary distribution of the different Plasmodium species as well as the anti-malarial drug profile. Results Of the 152 participants sampled, 98 (64.5%) were females. The average age of the participants was 34.9 years (range: 2 months–81 years). There were 134 samples that showed PCR amplification, confirming the species as Plasmodium. Plasmodium falciparum (N = 122), Plasmodium malariae (N = 6), Plasmodium ovale (N = 4), and Plasmodium vivax (N = 2) were the various Plasmodium species and their proportions. The results showed that 87 (71.3%) of the samples were sensitive strains/wild type (CVMNK), 4 (3.3%) were resistant haplotypes (SVMNT), and 31 (25.4%) were resistant haplotypes (CVIET). Kelch13 C580Y mutation was not detected. Conclusion The community served by Kisoro hospital has a high Plasmodium species burden, according to this study. Plasmodium falciparum was the dominant species, and it has shown that resistance to chloroquine is decreasing in the region. Based on this, molecular identification of Plasmodium species is critical for better clinical management. Besides, DBS is an appropriate medium for DNA preservation and storage for future epidemiological studies.

HRP2 and PLDH Performance Comparison as Rapid Diagnostic Tools for Symptomatic Malaria Patients at Rugazi HC IV in Rubirizi District

“…A study conducted in four contagious villages in Mulanda subcounty in Tororo District, Eastern Uganda, showed that infection due to Plasmodium falciparum was 94.0%, which was also the most common species identified; Plasmodium malariae was 6.0%, and there were no mixed infections seen [45]. Similar studies conducted in selected health care facilities across Ghana and Kisoro Hospital in southwestern Uganda showed results that were not significantly different from our study, with the exception of P. malariae, which was not identified in our study [24,46]. Studies in the same region of Uganda show results that are not significantly different from our study results [47][48][49].…”

Antimalarial Drug Resistance: A Brief History of Its Spread in Indonesia

“…Evaluation for drug resistance is highly recommended to determine the success of malaria treatment. 61 The identification of antimalarial drug resistance in Plasmodium species can be performed through several methods, including in vivo, drug level, in vitro, ex vivo, and molecular characterization. 62,63 In 2009, WHO established a protocol to detect antimalarial drug resistance.…”

“…Moreover, sequencing methods are performed using either Sanger or nextgeneration sequencing (NGS), such as Illumina MiSeq, Pyrosequencing, and MinION nanopore, which are suitable to identify numerous genetic markers for antimalarial drug resistance in a large number of samples for each run. 61,[67][68][69] Application of various detection methods in antimalarial drug resistance depends on the facility in malaria endemic areas. In Indonesia, TES is commonly used for the studies of drug resistant, 8,70,71 as well as molecular methods, such as PCR (nested and RFLP).…”

“…Moreover, sequencing methods are performed using either Sanger or next-generation sequencing (NGS), such as Illumina MiSeq, Pyrosequencing, and MinION nanopore, which are suitable to identify numerous genetic markers for antimalarial drug resistance in a large number of samples for each run. 61 , 67–69 …”

Diagnosis and management of malaria in the intensive care unit

“…[ [31] , [32] , [33] , [34] , [35] ] Although this initiative has impacted and reduced the global burden of the disease, malaria diagnosis, and management in the ICU remain a challenge amidst threats imposed by antimicrobial resistance. [ 19 , 36 , 37 ] This review provides valuable and critical information on the diagnosis and management of malaria in the ICU.…”