PCR detection of <it>Plasmodium falciparum </it>in human urine and saliva samples
<p>Abstract</p> <p>Background</p> <p>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 <it>Plasmodium fal...
| Main Authors: | , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
BMC
2006-11-01
|
| Series: | Malaria Journal |
| Online Access: | http://www.malariajournal.com/content/5/1/103 |
| _version_ | 1811266314608574464 |
|---|---|
| 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 |
| collection | DOAJ |
| description | <p>Abstract</p> <p>Background</p> <p>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 <it>Plasmodium falciparum </it>in human urine and saliva samples, and illustrates this potential application in genotyping malaria infections.</p> <p>Methods</p> <p>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. <it>P. falciparum </it>DNA was extracted from blood, urine and saliva, in separate groups, using the Chelex method or Qiagen DNEasy<sup>® </sup>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.</p> <p>Results and discussion</p> <p>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<sup>® </sup>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.</p> <p>Conclusion</p> <p><it>P. falciparum </it>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.</p> |
| first_indexed | 2024-04-12T20:41:01Z |
| format | Article |
| id | doaj.art-ee57d48c407b42cba2734ad443982289 |
| institution | Directory Open Access Journal |
| issn | 1475-2875 |
| language | English |
| last_indexed | 2024-04-12T20:41:01Z |
| publishDate | 2006-11-01 |
| publisher | BMC |
| record_format | Article |
| series | Malaria Journal |
| spelling | doaj.art-ee57d48c407b42cba2734ad4439822892022-12-22T03:17:26ZengBMCMalaria Journal1475-28752006-11-015110310.1186/1475-2875-5-103PCR detection of <it>Plasmodium falciparum </it>in human urine and saliva samplesThuma Philip EShiff Clive JSimoloka ChristopherMharakurwa SunganoSullivan David J<p>Abstract</p> <p>Background</p> <p>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 <it>Plasmodium falciparum </it>in human urine and saliva samples, and illustrates this potential application in genotyping malaria infections.</p> <p>Methods</p> <p>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. <it>P. falciparum </it>DNA was extracted from blood, urine and saliva, in separate groups, using the Chelex method or Qiagen DNEasy<sup>® </sup>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.</p> <p>Results and discussion</p> <p>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<sup>® </sup>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.</p> <p>Conclusion</p> <p><it>P. falciparum </it>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.</p>http://www.malariajournal.com/content/5/1/103 |
| spellingShingle | Thuma Philip E Shiff Clive J Simoloka Christopher Mharakurwa Sungano Sullivan David J PCR detection of <it>Plasmodium falciparum </it>in human urine and saliva samples Malaria Journal |
| title | PCR detection of <it>Plasmodium falciparum </it>in human urine and saliva samples |
| title_full | PCR detection of <it>Plasmodium falciparum </it>in human urine and saliva samples |
| title_fullStr | PCR detection of <it>Plasmodium falciparum </it>in human urine and saliva samples |
| title_full_unstemmed | PCR detection of <it>Plasmodium falciparum </it>in human urine and saliva samples |
| title_short | PCR detection of <it>Plasmodium falciparum </it>in human urine and saliva samples |
| title_sort | pcr detection of it plasmodium falciparum it in human urine and saliva samples |
| url | http://www.malariajournal.com/content/5/1/103 |
| work_keys_str_mv | AT thumaphilipe pcrdetectionofitplasmodiumfalciparumitinhumanurineandsalivasamples AT shiffclivej pcrdetectionofitplasmodiumfalciparumitinhumanurineandsalivasamples AT simolokachristopher pcrdetectionofitplasmodiumfalciparumitinhumanurineandsalivasamples AT mharakurwasungano pcrdetectionofitplasmodiumfalciparumitinhumanurineandsalivasamples AT sullivandavidj pcrdetectionofitplasmodiumfalciparumitinhumanurineandsalivasamples |