Apr 22, 2024

Public workspaceDNA extraction and detection of Mansonella perstans from Dry Blood Spots (DBS)

DOI
https://dx.doi.org/10.17504/protocols.io.x54v928b4l3e/v1
  • Millicent Opoku1,
  • Dziedzom De Souza2
  • 1La Trobe University, Australia;
  • 2Department of Parasitology, Noguchi Memorial Institute for Medical Research, University of Ghana, Ghana
Millicent Opoku
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DOI: https://dx.doi.org/10.17504/protocols.io.x54v928b4l3e/v1
Document Citation: Millicent Opoku, Dziedzom De Souza 2024. DNA extraction and detection of Mansonella perstans from Dry Blood Spots (DBS). protocols.io https://dx.doi.org/10.17504/protocols.io.x54v928b4l3e/v1
License: This is an open access document distributed under the terms of the Creative Commons Attribution License,  which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Created: April 04, 2024
Last Modified: April 22, 2024
Document Integer ID: 97770
Keywords: DNA, Dried Blood Spot, Mansonella, Nested PCR, Excised Gels, detection of mansonella perstan, mansonellosis in human, mansonella perstan, mansonellosi, dna extraction, dried blood blot, filarial parasite, dry blood spot, other filarial parasitic disease, blood blot, dna, blood, disease, extraction
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Abstract
Mansonella perstans is a filarial parasite that causes mansonellosis in humans. Unlike other filarial parasitic diseases, mansonellosis is rarely symptomatic and hence poorly studied. This protocol describes the detection of M. perstans in blood. Firstly, DNA was extracted from dried blood blots (DBS). The DNA were screened by a nested PCR and subsequently sanger sequencing was done on excised gels. The sequences were then compared to reference sequences deposited in NCBI.
Troubleshooting
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Protocol references
1. Quick-DNA‱ MiniPrep (Zymo Research, USA)
2.Tang T-HT, López-Vélez R, Lanza M, Shelley AJ, Rubio JM, Luz SLB. Nested PCR to detect and distinguish the sympatric filarial species Onchocerca volvulus, Mansonella ozzardi and Mansonella perstans in the Amazon Region. Memórias do Instituto Oswaldo Cruz. 2010;105(6):823-8.