Abstract:
Malaria elimination strategies demand constant monitoring of the parasite population for genetic variations that necessitate a public health response, such as a new form of drug resistance. Malaria control relies mainly on rapid and precise diagnosis, followed by successful treatment. Malaria diagnosis must be made as soon as possible to provide optimal disease management and surveillance. In every situation, the accuracy of malaria diagnosis is essential, as misdiagnosis can result in severe morbidity and mortality. Here, we describe a novel, high through put method using an Illumina Mi-Seq platform to demonstrate the proportions of Plasmodium species in meta barcoded DNA samples derived from human malaria patients. We reported a unique, high throughput approach for determining the ratio of Plasmodium species in meta barcoding DNA samples generated from human malaria patients using an Illumina MiSeq. Positive control gDNA from P. falciparum and P. vivax was used to mock DNA pols of parasites to test the assay direction threshold for each species. Several mock pools indicated the accuracy of detection abilities and the proportion of each species. The technique was subsequently used on malaria-positive patient samples to determine the species composition of Plasmodium populations in Punjab region from Pakistan and the tribal territories of the Auragzai Agency Border (Shanawari Zargari), Ali Masjid, Landi-Kotal Khyber Agency on Pak-Afghan border. The deep amplicon sequencing approach contrasts with an immunochromatographic test, commonly utilized for diagnosis in the region. According to deep amplicon sequencing, P. vivax was present in 69.8 percent of the patients, P. falciparum in 29.5 percent and mixed infection in 0.7 percent of the cases. Plasmodium vivax was found in 65.6 percent of patients, P. falciparum in 27.4 percent, mixed infection in 0.7 percent of patients and 6.32 percent of positive malaria cases were negative in immunochromatographic diagnosis but positive in deep amplicon sequencing. Overall, metabarcoding DNA sequencing improves diagnosis accuracy, resulting in a significant increase in Plasmodium infection prevalence estimates. The use of metabarcoding DNA in next-generation sequencing could help in Plasmodium infection diagnosis, surveillance, treatment, and control and can also contribute to research in parasite biology.
Page(s):
0-0
DOI:
DOI not available
Published:
Journal: First International Conference on Revamped Scientific Outlook of 21st Century (Abstract Book), Volume: 0, Issue: 0, Year: 2022
Keywords:
Plasmodium
,
diagnosis
,
Surveillance
,
Surveillance
,
Amplicon sequencing
,
Surveillance
,
Surveillance
,
P falciparum
,
Immunechromatographic
,
Illumina