Filter paper collection of Plasmodium falciparum mRNA for detecting low-density gametocytes
| Autor: | Jones Sophie, Sutherland Colin J, Hermsen Cornelus, Arens Theo, Teelen Karina, Hallett Rachel, Corran Patrick, van der Vegte-Bolmer Marga, Sauerwein Robert, Drakeley Chris J, Bousema Teun |
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| Jazyk: | angličtina |
| Rok vydání: | 2012 |
| Předmět: |
Reverse-transcription polymerase chain reaction (RT-PCR)
Quantitative nucleic acid sequence-based amplification (QT-NASBA) Sub-microscopic Gametocyte Detection Elimination Transmission Ribonucleic acid (RNA) Filter paper Arctic medicine. Tropical medicine RC955-962 Infectious and parasitic diseases RC109-216 |
| Zdroj: | Malaria Journal, Vol 11, Iss 1, p 266 (2012) |
| Druh dokumentu: | article |
| ISSN: | 1475-2875 |
| DOI: | 10.1186/1475-2875-11-266 |
| Popis: | Abstract Background Accurate sampling of sub-microscopic gametocytes is necessary for epidemiological studies to identify the infectious reservoir of Plasmodium falciparum. Detection of gametocyte mRNA achieves sensitive detection, but requires careful handling of samples. Filter papers can be used for collecting RNA samples, but rigorous testing of their capacity to withstand adverse storage conditions has not been fully explored. Methods Three gametocyte dilutions: 10/μL, 1.0/μL and 0.1/μL were spotted onto Whatman™ 903 Protein Saver Cards, FTA Classic Cards and 3MM filter papers that were stored under frozen, cold chain or tropical conditions for up to 13 weeks . RNA was extracted, then detected by quantitative nucleic acid sequence-based amplification (QT-NASBA) and reverse-transcriptase PCR (RT-PCR). Results Successful gametocyte detection was more frequently observed from the Whatman 903 Protein Saver Card compared to the Whatman FTA Classic Card, by both techniques (p Conclusions This study indicates the Whatman 903 Protein Saver Card is better for Pfs25 mRNA sampling compared to the Whatman FTA Classic Card, and that the Whatman 3MM filter paper may prove to be a satisfactory cheaper option for Pfs25 mRNA sampling. When appropriately dried, filter papers provide a useful approach to Pfs25 mRNA sampling, especially in settings where storage in RNA-protecting buffer is not possible. |
| Databáze: | Directory of Open Access Journals |
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