Molecular Detection of Plasmodium vivax in Stored Blood Smears using Nested PCR
DOI:
10.29303/jbt.v26i4.12376Published:
2026-09-22Downloads
Abstract
Malaria is an infectious disease caused by Plasmodium parasites and transmitted by female Anopheles mosquitoes. Plasmodium vivax is widely distributed and can form hypnozoites that cause relapse. Detection may be difficult, particularly in samples with low parasite density. This descriptive study used 23 stored Giemsa stained blood smear samples previously confirmed as P. vivax by microscopic examination. Positive and negative controls were included to monitor the amplification process. Agarose gel electrophoresis was used to visualize the PCR products. No expected bands were detected in the first or second amplification stages. In the third stage, 10 of 23 samples (43.48%) produced the expected 121 bp band and were identified as positive for P. vivax. The absence of expected bands in the earlier stages may indicate DNA fragmentation caused by methanol fixation, Giemsa staining, and prolonged storage. The main limitation of this study was the use of stored samples with potentially degraded DNA, which may have affected amplification results. These findings indicate that Nested PCR with species-specific primers can detect P. vivax DNA in stored blood smear samples, particularly when short DNA fragments remain available for amplification.
Keywords:
Plasmodium vivax, Stored Blood Smear, Nested PCR, Molecular Detection, Malaria DiagnosticsReferences
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