A dual colour fluorescence in situ hybridization (FISH) assay for identifying the zoonotic malaria parasite Plasmodium knowlesi with a potential application for the specific diagnosis of knowlesi malaria in peripheral-level laboratories of Southeast Asia
Abstract Background Plasmodium knowlesi is primarily responsible for zoonotic malaria in several Southeast Asian countries. Precise identification of the parasite in the blood of patients presently relies on an expensive and elaborate PCR procedure because microscopic examination of blood and other...
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| Format: | Article |
| Language: | English |
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BMC
2017-07-01
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| Series: | Parasites & Vectors |
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| Online Access: | http://link.springer.com/article/10.1186/s13071-017-2273-7 |
| _version_ | 1811260186148470784 |
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| author | Jyotsna Shah Akhila Poruri Olivia Mark Urmila Khadilka Franziska Mohring Robert W. Moon Ranjan Ramasamy |
| author_facet | Jyotsna Shah Akhila Poruri Olivia Mark Urmila Khadilka Franziska Mohring Robert W. Moon Ranjan Ramasamy |
| author_sort | Jyotsna Shah |
| collection | DOAJ |
| description | Abstract Background Plasmodium knowlesi is primarily responsible for zoonotic malaria in several Southeast Asian countries. Precise identification of the parasite in the blood of patients presently relies on an expensive and elaborate PCR procedure because microscopic examination of blood and other available field identification techniques lack adequate specificity. Therefore, the use of a simple and inexpensive dual-colour fluorescence in situ hybridization (FISH) assay, analogous to FISH assays recently described for Plasmodium falciparum and Plasmodium vivax, was investigated as a potential tool for identifying P. knowlesi. Results A P. knowlesi 18S rDNA sequence-based DNA probe was used to test thin blood smears of P. knowlesi by FISH, and fluorescence viewed in a light microscope fitted with a light emitting diode light source and appropriate emission and barrier filters. The limit of detection in the P. knowlesi FISH assay was 84 parasites per μl in infected monkey blood and 61 parasites per μl for P. knowlesi cultured in human blood. The P. knowlesi-specific FISH probe detected only P. knowlesi and not P. falciparum, Plasmodium malariae, Plasmodium ovale, P. vivax or a panel of other human blood-borne pathogens. A previously described Plasmodium genus-specific probe used simultaneously in the P. knowlesi FISH assay reacted with all tested Plasmodium species. Conclusions To our knowledge, this is the first description of a FISH assay for P. knowlesi that is potentially useful for diagnosing infections in remote laboratories in endemic countries. |
| first_indexed | 2024-04-12T18:43:43Z |
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| id | doaj.art-a52d5a0824bc4e41a3f7d2506c43d59a |
| institution | Directory Open Access Journal |
| issn | 1756-3305 |
| language | English |
| last_indexed | 2024-04-12T18:43:43Z |
| publishDate | 2017-07-01 |
| publisher | BMC |
| record_format | Article |
| series | Parasites & Vectors |
| spelling | doaj.art-a52d5a0824bc4e41a3f7d2506c43d59a2022-12-22T03:20:42ZengBMCParasites & Vectors1756-33052017-07-011011910.1186/s13071-017-2273-7A dual colour fluorescence in situ hybridization (FISH) assay for identifying the zoonotic malaria parasite Plasmodium knowlesi with a potential application for the specific diagnosis of knowlesi malaria in peripheral-level laboratories of Southeast AsiaJyotsna Shah0Akhila Poruri1Olivia Mark2Urmila Khadilka3Franziska Mohring4Robert W. Moon5Ranjan Ramasamy6ID-FISH TechnologyID-FISH TechnologyID-FISH TechnologyKasturba Medical College HospitalLondon School of Hygiene and Tropical MedicineLondon School of Hygiene and Tropical MedicineID-FISH TechnologyAbstract Background Plasmodium knowlesi is primarily responsible for zoonotic malaria in several Southeast Asian countries. Precise identification of the parasite in the blood of patients presently relies on an expensive and elaborate PCR procedure because microscopic examination of blood and other available field identification techniques lack adequate specificity. Therefore, the use of a simple and inexpensive dual-colour fluorescence in situ hybridization (FISH) assay, analogous to FISH assays recently described for Plasmodium falciparum and Plasmodium vivax, was investigated as a potential tool for identifying P. knowlesi. Results A P. knowlesi 18S rDNA sequence-based DNA probe was used to test thin blood smears of P. knowlesi by FISH, and fluorescence viewed in a light microscope fitted with a light emitting diode light source and appropriate emission and barrier filters. The limit of detection in the P. knowlesi FISH assay was 84 parasites per μl in infected monkey blood and 61 parasites per μl for P. knowlesi cultured in human blood. The P. knowlesi-specific FISH probe detected only P. knowlesi and not P. falciparum, Plasmodium malariae, Plasmodium ovale, P. vivax or a panel of other human blood-borne pathogens. A previously described Plasmodium genus-specific probe used simultaneously in the P. knowlesi FISH assay reacted with all tested Plasmodium species. Conclusions To our knowledge, this is the first description of a FISH assay for P. knowlesi that is potentially useful for diagnosing infections in remote laboratories in endemic countries.http://link.springer.com/article/10.1186/s13071-017-2273-7DNA probeFluorescence in situ hybridizationMalaria diagnosisPlasmodium knowlesiZoonotic malaria |
| spellingShingle | Jyotsna Shah Akhila Poruri Olivia Mark Urmila Khadilka Franziska Mohring Robert W. Moon Ranjan Ramasamy A dual colour fluorescence in situ hybridization (FISH) assay for identifying the zoonotic malaria parasite Plasmodium knowlesi with a potential application for the specific diagnosis of knowlesi malaria in peripheral-level laboratories of Southeast Asia Parasites & Vectors DNA probe Fluorescence in situ hybridization Malaria diagnosis Plasmodium knowlesi Zoonotic malaria |
| title | A dual colour fluorescence in situ hybridization (FISH) assay for identifying the zoonotic malaria parasite Plasmodium knowlesi with a potential application for the specific diagnosis of knowlesi malaria in peripheral-level laboratories of Southeast Asia |
| title_full | A dual colour fluorescence in situ hybridization (FISH) assay for identifying the zoonotic malaria parasite Plasmodium knowlesi with a potential application for the specific diagnosis of knowlesi malaria in peripheral-level laboratories of Southeast Asia |
| title_fullStr | A dual colour fluorescence in situ hybridization (FISH) assay for identifying the zoonotic malaria parasite Plasmodium knowlesi with a potential application for the specific diagnosis of knowlesi malaria in peripheral-level laboratories of Southeast Asia |
| title_full_unstemmed | A dual colour fluorescence in situ hybridization (FISH) assay for identifying the zoonotic malaria parasite Plasmodium knowlesi with a potential application for the specific diagnosis of knowlesi malaria in peripheral-level laboratories of Southeast Asia |
| title_short | A dual colour fluorescence in situ hybridization (FISH) assay for identifying the zoonotic malaria parasite Plasmodium knowlesi with a potential application for the specific diagnosis of knowlesi malaria in peripheral-level laboratories of Southeast Asia |
| title_sort | dual colour fluorescence in situ hybridization fish assay for identifying the zoonotic malaria parasite plasmodium knowlesi with a potential application for the specific diagnosis of knowlesi malaria in peripheral level laboratories of southeast asia |
| topic | DNA probe Fluorescence in situ hybridization Malaria diagnosis Plasmodium knowlesi Zoonotic malaria |
| url | http://link.springer.com/article/10.1186/s13071-017-2273-7 |
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