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Microfilaria Positification Test Using Real-Time PCR Technique with HRM (High-Resolution Melting)
Bagus Muhammad Ihsan1, Afiat Berbudi2, Ridad Agoes2
1Graduate School of Biomedical Sciences Master Program, Faculty of Medicine, Universitas Padjadjaran, Bandung, Indonesia.
Acta Parasitologica
|June 17, 2021
Summary
Microscopic examination often misses filariasis microfilaria. Quantitative Polymerase Chain Reaction with High-Resolution Melting (qPCR-HRM) effectively identifies Brugia malayi and Wuchereria bancrofti in patient samples.
Area of Science:
- Molecular Biology
- Parasitology
- Medical Diagnostics
Background:
- Microscopic examination for filariasis diagnosis has limitations in detecting microfilaria.
- Polymerase Chain Reaction (PCR) methods, including High-Resolution Melting (HRM), offer alternative diagnostic approaches.
Purpose of the Study:
- To compare the efficacy of quantitative PCR with High-Resolution Melting (qPCR-HRM) against traditional microscopic examination for identifying filarial worm species.
Main Methods:
- 19 patient samples were analyzed using both standard microscopy and qPCR-HRM.
- qPCR-HRM utilized specific melting temperatures to differentiate microfilaria types.
Main Results:
- Microscopic examination failed to detect any microfilaria in the analyzed samples.
- qPCR-HRM successfully identified Brugia malayi and Wuchereria bancrofti, with distinct melting temperatures of 78.1–78.7°C and 80.2–80.8°C, respectively.
Conclusions:
- qPCR-HRM is a more sensitive method than microscopy for detecting and identifying W. bancrofti and B. malayi.
- This molecular technique enhances filariasis diagnosis by accurately identifying causative agents.
