PCR detection of malaria parasites in desiccated Anopheles mosquitoes is uninhibited by storage time and temperature
<p><b>Abstract</b></p> <p><b>Background</b></p> <p>Reliable methods to preserve mosquito vectors for malaria studies are necessary for detecting <it>Plasmodium</it> parasites. In field settings, however, maintaining a cold chain of st...
| Main Authors: | , , , , |
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| Format: | Article |
| Language: | English |
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BMC
2012-06-01
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| Series: | Malaria Journal |
| Subjects: | |
| Online Access: | http://www.malariajournal.com/content/11/1/193 |
| _version_ | 1819043994533888000 |
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| author | Rider Mark A Byrd Brian D Keating Joseph Wesson Dawn M Caillouet Kevin A |
| author_facet | Rider Mark A Byrd Brian D Keating Joseph Wesson Dawn M Caillouet Kevin A |
| author_sort | Rider Mark A |
| collection | DOAJ |
| description | <p><b>Abstract</b></p> <p><b>Background</b></p> <p>Reliable methods to preserve mosquito vectors for malaria studies are necessary for detecting <it>Plasmodium</it> parasites. In field settings, however, maintaining a cold chain of storage from the time of collection until laboratory processing, or accessing other reliable means of sample preservation is often logistically impractical or cost prohibitive. As the <it>Plasmodium</it> infection rate of <it>Anopheles</it> mosquitoes is a central component of the entomological inoculation rate and other indicators of transmission intensity, storage conditions that affect pathogen detection may bias malaria surveillance indicators. This study investigated the effect of storage time and temperature on the ability to detect <it>Plasmodium</it> parasites in desiccated <it>Anopheles</it> mosquitoes by real-time polymerase chain reaction (PCR).</p> <p><b>Methods</b></p> <p>Laboratory-infected <it>Anopheles stephensi</it> mosquitoes were chloroform-killed and stored over desiccant for 0, 1, 3, and 6 months while being held at four different temperatures: 28, 37, -20 and -80°C. The detection of <it>Plasmodium</it> DNA was evaluated by real-time PCR amplification of a 111 base pair region of block 4 of the merozoite surface protein.</p> <p><b>Results</b></p> <p>Varying the storage time and temperature of desiccated mosquitoes did not impact the sensitivity of parasite detection. A two-way factorial analysis of variance suggested that storage time and temperature were not associated with a loss in the ability to detect parasites. Storage of samples at 28°C resulted in a significant increase in the ability to detect parasite DNA, though no other positive associations were observed between the experimental storage treatments and PCR amplification.</p> <p><b>Conclusions</b></p> <p>Cold chain maintenance of desiccated mosquito samples is not necessary for real-time PCR detection of parasite DNA. Though field-collected mosquitoes may be subjected to variable conditions prior to molecular processing, the storage of samples over an inexpensive and logistically accessible desiccant will likely ensure accurate assessment of malaria parasite presence without diminishing PCR-detection of parasites in mosquitoes stored for at least six months.</p> |
| first_indexed | 2024-12-21T10:05:37Z |
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| id | doaj.art-1b1f96ec53ce464a965f89f91cda29a9 |
| institution | Directory Open Access Journal |
| issn | 1475-2875 |
| language | English |
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| spelling | doaj.art-1b1f96ec53ce464a965f89f91cda29a92022-12-21T19:07:50ZengBMCMalaria Journal1475-28752012-06-0111119310.1186/1475-2875-11-193PCR detection of malaria parasites in desiccated Anopheles mosquitoes is uninhibited by storage time and temperatureRider Mark AByrd Brian DKeating JosephWesson Dawn MCaillouet Kevin A<p><b>Abstract</b></p> <p><b>Background</b></p> <p>Reliable methods to preserve mosquito vectors for malaria studies are necessary for detecting <it>Plasmodium</it> parasites. In field settings, however, maintaining a cold chain of storage from the time of collection until laboratory processing, or accessing other reliable means of sample preservation is often logistically impractical or cost prohibitive. As the <it>Plasmodium</it> infection rate of <it>Anopheles</it> mosquitoes is a central component of the entomological inoculation rate and other indicators of transmission intensity, storage conditions that affect pathogen detection may bias malaria surveillance indicators. This study investigated the effect of storage time and temperature on the ability to detect <it>Plasmodium</it> parasites in desiccated <it>Anopheles</it> mosquitoes by real-time polymerase chain reaction (PCR).</p> <p><b>Methods</b></p> <p>Laboratory-infected <it>Anopheles stephensi</it> mosquitoes were chloroform-killed and stored over desiccant for 0, 1, 3, and 6 months while being held at four different temperatures: 28, 37, -20 and -80°C. The detection of <it>Plasmodium</it> DNA was evaluated by real-time PCR amplification of a 111 base pair region of block 4 of the merozoite surface protein.</p> <p><b>Results</b></p> <p>Varying the storage time and temperature of desiccated mosquitoes did not impact the sensitivity of parasite detection. A two-way factorial analysis of variance suggested that storage time and temperature were not associated with a loss in the ability to detect parasites. Storage of samples at 28°C resulted in a significant increase in the ability to detect parasite DNA, though no other positive associations were observed between the experimental storage treatments and PCR amplification.</p> <p><b>Conclusions</b></p> <p>Cold chain maintenance of desiccated mosquito samples is not necessary for real-time PCR detection of parasite DNA. Though field-collected mosquitoes may be subjected to variable conditions prior to molecular processing, the storage of samples over an inexpensive and logistically accessible desiccant will likely ensure accurate assessment of malaria parasite presence without diminishing PCR-detection of parasites in mosquitoes stored for at least six months.</p>http://www.malariajournal.com/content/11/1/193<it>Plasmodium</it> parasite detectionDNATemperatureDesicant<it>Anopheles</it>Storage |
| spellingShingle | Rider Mark A Byrd Brian D Keating Joseph Wesson Dawn M Caillouet Kevin A PCR detection of malaria parasites in desiccated Anopheles mosquitoes is uninhibited by storage time and temperature Malaria Journal <it>Plasmodium</it> parasite detection DNA Temperature Desicant <it>Anopheles</it> Storage |
| title | PCR detection of malaria parasites in desiccated Anopheles mosquitoes is uninhibited by storage time and temperature |
| title_full | PCR detection of malaria parasites in desiccated Anopheles mosquitoes is uninhibited by storage time and temperature |
| title_fullStr | PCR detection of malaria parasites in desiccated Anopheles mosquitoes is uninhibited by storage time and temperature |
| title_full_unstemmed | PCR detection of malaria parasites in desiccated Anopheles mosquitoes is uninhibited by storage time and temperature |
| title_short | PCR detection of malaria parasites in desiccated Anopheles mosquitoes is uninhibited by storage time and temperature |
| title_sort | pcr detection of malaria parasites in desiccated anopheles mosquitoes is uninhibited by storage time and temperature |
| topic | <it>Plasmodium</it> parasite detection DNA Temperature Desicant <it>Anopheles</it> Storage |
| url | http://www.malariajournal.com/content/11/1/193 |
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