PCR detection of Plasmodium falciparum in human urine and saliva samples
Abstract Background Current detection or screening for malaria infection necessitates drawing blood by fingerprick or venipuncture, which poses risks and limitations for repeated measurement. This study presents PCR detection of Plasmodium falciparum in human urine and saliva samples, and illustrate...
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ftdoajarticles:oai:doaj.org/article:ee57d48c407b42cba2734ad443982289 2023-05-15T15:16:52+02:00 PCR detection of Plasmodium falciparum in human urine and saliva samples Thuma Philip E Shiff Clive J Simoloka Christopher Mharakurwa Sungano Sullivan David J 2006-11-01T00:00:00Z https://doi.org/10.1186/1475-2875-5-103 https://doaj.org/article/ee57d48c407b42cba2734ad443982289 EN eng BMC http://www.malariajournal.com/content/5/1/103 https://doaj.org/toc/1475-2875 doi:10.1186/1475-2875-5-103 1475-2875 https://doaj.org/article/ee57d48c407b42cba2734ad443982289 Malaria Journal, Vol 5, Iss 1, p 103 (2006) Arctic medicine. Tropical medicine RC955-962 Infectious and parasitic diseases RC109-216 article 2006 ftdoajarticles https://doi.org/10.1186/1475-2875-5-103 2022-12-30T21:57:52Z Abstract Background Current detection or screening for malaria infection necessitates drawing blood by fingerprick or venipuncture, which poses risks and limitations for repeated measurement. This study presents PCR detection of Plasmodium falciparum in human urine and saliva samples, and illustrates this potential application in genotyping malaria infections. Methods Urine and saliva were obtained from 47 thick film positive and 4 negative individuals one day after collection of blood slides and filter paper blood spots. P. falciparum DNA was extracted from blood, urine and saliva, in separate groups, using the Chelex method or Qiagen DNEasy ® kit (urine and saliva only). Blood, urine and saliva extracts were subjected to PCR in separate batches. Amplicons from the various sample types were examined for MSP2 polymorphisms and restriction fragment patterns on DHFR amino acid codon 59. Results and discussion Malaria infections exhibited primarily low-grade parasite densities, with a geometric mean of 775 asexual parasites/μl. Regularly matching polymorphic MSP2 genotypes were found between the corresponding urine, saliva and peripheral blood amplicons of each individual, with different inter-individual polymorphic genotypes. Amplicon yields were significantly dependent on DNA extraction method, parasite density and primer set (p < 0.001). A Qiagen ® kit extraction had more than 2× higher amplicon yield than the Chelex method, for both urine and saliva. Amplicon yields were 1.6 fold higher from saliva than urine. For each unit increase in log parasite density, the probability of amplicon enhanced 1.8 fold. Highest amplicon yields were obtained from the primer set with the shortest PCR product. Conclusion P. falciparum infection is detectable by PCR on human urine and saliva samples. Subject to further refinement of extraction technique and amplicon yields, large-scale malaria parasite screening and epidemiological surveys could be possible without the need to collect blood and use of needles or sharps. Article in Journal/Newspaper Arctic Directory of Open Access Journals: DOAJ Articles Arctic Malaria Journal 5 1 103 |
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Directory of Open Access Journals: DOAJ Articles |
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Arctic medicine. Tropical medicine RC955-962 Infectious and parasitic diseases RC109-216 |
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Arctic medicine. Tropical medicine RC955-962 Infectious and parasitic diseases RC109-216 Thuma Philip E Shiff Clive J Simoloka Christopher Mharakurwa Sungano Sullivan David J PCR detection of Plasmodium falciparum in human urine and saliva samples |
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Arctic medicine. Tropical medicine RC955-962 Infectious and parasitic diseases RC109-216 |
description |
Abstract Background Current detection or screening for malaria infection necessitates drawing blood by fingerprick or venipuncture, which poses risks and limitations for repeated measurement. This study presents PCR detection of Plasmodium falciparum in human urine and saliva samples, and illustrates this potential application in genotyping malaria infections. Methods Urine and saliva were obtained from 47 thick film positive and 4 negative individuals one day after collection of blood slides and filter paper blood spots. P. falciparum DNA was extracted from blood, urine and saliva, in separate groups, using the Chelex method or Qiagen DNEasy ® kit (urine and saliva only). Blood, urine and saliva extracts were subjected to PCR in separate batches. Amplicons from the various sample types were examined for MSP2 polymorphisms and restriction fragment patterns on DHFR amino acid codon 59. Results and discussion Malaria infections exhibited primarily low-grade parasite densities, with a geometric mean of 775 asexual parasites/μl. Regularly matching polymorphic MSP2 genotypes were found between the corresponding urine, saliva and peripheral blood amplicons of each individual, with different inter-individual polymorphic genotypes. Amplicon yields were significantly dependent on DNA extraction method, parasite density and primer set (p < 0.001). A Qiagen ® kit extraction had more than 2× higher amplicon yield than the Chelex method, for both urine and saliva. Amplicon yields were 1.6 fold higher from saliva than urine. For each unit increase in log parasite density, the probability of amplicon enhanced 1.8 fold. Highest amplicon yields were obtained from the primer set with the shortest PCR product. Conclusion P. falciparum infection is detectable by PCR on human urine and saliva samples. Subject to further refinement of extraction technique and amplicon yields, large-scale malaria parasite screening and epidemiological surveys could be possible without the need to collect blood and use of needles or sharps. |
format |
Article in Journal/Newspaper |
author |
Thuma Philip E Shiff Clive J Simoloka Christopher Mharakurwa Sungano Sullivan David J |
author_facet |
Thuma Philip E Shiff Clive J Simoloka Christopher Mharakurwa Sungano Sullivan David J |
author_sort |
Thuma Philip E |
title |
PCR detection of Plasmodium falciparum in human urine and saliva samples |
title_short |
PCR detection of Plasmodium falciparum in human urine and saliva samples |
title_full |
PCR detection of Plasmodium falciparum in human urine and saliva samples |
title_fullStr |
PCR detection of Plasmodium falciparum in human urine and saliva samples |
title_full_unstemmed |
PCR detection of Plasmodium falciparum in human urine and saliva samples |
title_sort |
pcr detection of plasmodium falciparum in human urine and saliva samples |
publisher |
BMC |
publishDate |
2006 |
url |
https://doi.org/10.1186/1475-2875-5-103 https://doaj.org/article/ee57d48c407b42cba2734ad443982289 |
geographic |
Arctic |
geographic_facet |
Arctic |
genre |
Arctic |
genre_facet |
Arctic |
op_source |
Malaria Journal, Vol 5, Iss 1, p 103 (2006) |
op_relation |
http://www.malariajournal.com/content/5/1/103 https://doaj.org/toc/1475-2875 doi:10.1186/1475-2875-5-103 1475-2875 https://doaj.org/article/ee57d48c407b42cba2734ad443982289 |
op_doi |
https://doi.org/10.1186/1475-2875-5-103 |
container_title |
Malaria Journal |
container_volume |
5 |
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1 |
container_start_page |
103 |
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1766347167244484608 |