PCR detection of malaria parasites in desiccated Anopheles mosquitoes is uninhibited by storage time and temperature
Abstract Background Reliable methods to preserve mosquito vectors for malaria studies are necessary for detecting Plasmodium 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...
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ftdoajarticles:oai:doaj.org/article:1b1f96ec53ce464a965f89f91cda29a9 2023-05-15T15:14:59+02:00 PCR detection of malaria parasites in desiccated Anopheles mosquitoes is uninhibited by storage time and temperature Rider Mark A Byrd Brian D Keating Joseph Wesson Dawn M Caillouet Kevin A 2012-06-01T00:00:00Z https://doi.org/10.1186/1475-2875-11-193 https://doaj.org/article/1b1f96ec53ce464a965f89f91cda29a9 EN eng BMC http://www.malariajournal.com/content/11/1/193 https://doaj.org/toc/1475-2875 doi:10.1186/1475-2875-11-193 1475-2875 https://doaj.org/article/1b1f96ec53ce464a965f89f91cda29a9 Malaria Journal, Vol 11, Iss 1, p 193 (2012) Plasmodium parasite detection DNA Temperature Desicant Anopheles Storage Arctic medicine. Tropical medicine RC955-962 Infectious and parasitic diseases RC109-216 article 2012 ftdoajarticles https://doi.org/10.1186/1475-2875-11-193 2022-12-31T11:57:37Z Abstract Background Reliable methods to preserve mosquito vectors for malaria studies are necessary for detecting Plasmodium 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 Plasmodium infection rate of Anopheles 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 Plasmodium parasites in desiccated Anopheles mosquitoes by real-time polymerase chain reaction (PCR). Methods Laboratory-infected Anopheles stephensi 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 Plasmodium DNA was evaluated by real-time PCR amplification of a 111 base pair region of block 4 of the merozoite surface protein. Results 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. Conclusions 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 ... Article in Journal/Newspaper Arctic Directory of Open Access Journals: DOAJ Articles Arctic Malaria Journal 11 1 |
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topic |
Plasmodium parasite detection DNA Temperature Desicant Anopheles Storage Arctic medicine. Tropical medicine RC955-962 Infectious and parasitic diseases RC109-216 |
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Plasmodium parasite detection DNA Temperature Desicant Anopheles Storage Arctic medicine. Tropical medicine RC955-962 Infectious and parasitic diseases RC109-216 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 |
topic_facet |
Plasmodium parasite detection DNA Temperature Desicant Anopheles Storage Arctic medicine. Tropical medicine RC955-962 Infectious and parasitic diseases RC109-216 |
description |
Abstract Background Reliable methods to preserve mosquito vectors for malaria studies are necessary for detecting Plasmodium 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 Plasmodium infection rate of Anopheles 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 Plasmodium parasites in desiccated Anopheles mosquitoes by real-time polymerase chain reaction (PCR). Methods Laboratory-infected Anopheles stephensi 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 Plasmodium DNA was evaluated by real-time PCR amplification of a 111 base pair region of block 4 of the merozoite surface protein. Results 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. Conclusions 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 ... |
format |
Article in Journal/Newspaper |
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 |
title |
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_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_sort |
pcr detection of malaria parasites in desiccated anopheles mosquitoes is uninhibited by storage time and temperature |
publisher |
BMC |
publishDate |
2012 |
url |
https://doi.org/10.1186/1475-2875-11-193 https://doaj.org/article/1b1f96ec53ce464a965f89f91cda29a9 |
geographic |
Arctic |
geographic_facet |
Arctic |
genre |
Arctic |
genre_facet |
Arctic |
op_source |
Malaria Journal, Vol 11, Iss 1, p 193 (2012) |
op_relation |
http://www.malariajournal.com/content/11/1/193 https://doaj.org/toc/1475-2875 doi:10.1186/1475-2875-11-193 1475-2875 https://doaj.org/article/1b1f96ec53ce464a965f89f91cda29a9 |
op_doi |
https://doi.org/10.1186/1475-2875-11-193 |
container_title |
Malaria Journal |
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11 |
container_issue |
1 |
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1766345377085128704 |