Comparative evaluation of two commercial real-time PCR kits (QuantiFast™ and abTES™) for the detection of Plasmodium knowlesi and other Plasmodium species in Sabah, Malaysia
Abstract Background The monkey parasite Plasmodium knowlesi is an emerging public health issue in Southeast Asia. In Sabah, Malaysia, P. knowlesi is now the dominant cause of human malaria. Molecular detection methods for P. knowlesi are essential for accurate diagnosis and in monitoring progress to...
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| Format: | Article |
| Language: | English |
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BMC
2020-08-01
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| Series: | Malaria Journal |
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| Online Access: | http://link.springer.com/article/10.1186/s12936-020-03379-2 |
| _version_ | 1818112987694628864 |
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| author | Nor Afizah Nuin Angelica F. Tan Yao Long Lew Kim A. Piera Timothy William Giri S. Rajahram Jenarun Jelip Jiloris F. Dony Rashidah Mohammad Daniel J. Cooper Bridget E. Barber Nicholas M. Anstey Tock H. Chua Matthew J. Grigg |
| author_facet | Nor Afizah Nuin Angelica F. Tan Yao Long Lew Kim A. Piera Timothy William Giri S. Rajahram Jenarun Jelip Jiloris F. Dony Rashidah Mohammad Daniel J. Cooper Bridget E. Barber Nicholas M. Anstey Tock H. Chua Matthew J. Grigg |
| author_sort | Nor Afizah Nuin |
| collection | DOAJ |
| description | Abstract Background The monkey parasite Plasmodium knowlesi is an emerging public health issue in Southeast Asia. In Sabah, Malaysia, P. knowlesi is now the dominant cause of human malaria. Molecular detection methods for P. knowlesi are essential for accurate diagnosis and in monitoring progress towards malaria elimination of other Plasmodium species. However, recent commercially available PCR malaria kits have unpublished P. knowlesi gene targets or have not been evaluated against clinical samples. Methods Two real-time PCR methods currently used in Sabah for confirmatory malaria diagnosis and surveillance reporting were evaluated: the QuantiFast™ Multiplex PCR kit (Qiagen, Germany) targeting the P. knowlesi 18S SSU rRNA; and the abTES™ Malaria 5 qPCR II kit (AITbiotech, Singapore), with an undisclosed P. knowlesi gene target. Diagnostic accuracy was evaluated using 52 P. knowlesi, 25 Plasmodium vivax, 21 Plasmodium falciparum, and 10 Plasmodium malariae clinical isolates, and 26 malaria negative controls, and compared against a validated reference nested PCR assay. The limit of detection (LOD) for each PCR method and Plasmodium species was also evaluated. Results The sensitivity of the QuantiFast™ and abTES™ assays for detecting P. knowlesi was comparable at 98.1% (95% CI 89.7–100) and 100% (95% CI 93.2–100), respectively. Specificity of the QuantiFast™ and abTES™ for P. knowlesi was high at 98.8% (95% CI 93.4–100) for both assays. The QuantiFast™ assay demonstrated falsely-positive mixed Plasmodium species at low parasitaemias in both the primary and LOD analysis. Diagnostic accuracy of both PCR kits for detecting P. vivax, P. falciparum, and P. malariae was comparable to P. knowlesi. The abTES™ assay demonstrated a lower LOD for P. knowlesi of ≤ 0.125 parasites/µL compared to QuantiFast™ with a LOD of 20 parasites/µL. Hospital microscopy demonstrated a sensitivity of 78.8% (95% CI 65.3–88.9) and specificity of 80.4% (95% CI 67.6–89.8) compared to reference PCR for detecting P. knowlesi. Conclusion The QuantiFast™ and abTES™ commercial PCR kits performed well for the accurate detection of P. knowlesi infections. Although the QuantiFast™ kit is cheaper, the abTES™ kit demonstrated a lower LOD, supporting its use as a second-line referral-laboratory diagnostic tool in Sabah, Malaysia. |
| first_indexed | 2024-12-11T03:27:40Z |
| format | Article |
| id | doaj.art-5cf3067ff8904500a984a978891fc686 |
| institution | Directory Open Access Journal |
| issn | 1475-2875 |
| language | English |
| last_indexed | 2024-12-11T03:27:40Z |
| publishDate | 2020-08-01 |
| publisher | BMC |
| record_format | Article |
| series | Malaria Journal |
| spelling | doaj.art-5cf3067ff8904500a984a978891fc6862022-12-22T01:22:27ZengBMCMalaria Journal1475-28752020-08-0119111110.1186/s12936-020-03379-2Comparative evaluation of two commercial real-time PCR kits (QuantiFast™ and abTES™) for the detection of Plasmodium knowlesi and other Plasmodium species in Sabah, MalaysiaNor Afizah Nuin0Angelica F. Tan1Yao Long Lew2Kim A. Piera3Timothy William4Giri S. Rajahram5Jenarun Jelip6Jiloris F. Dony7Rashidah Mohammad8Daniel J. Cooper9Bridget E. Barber10Nicholas M. Anstey11Tock H. Chua12Matthew J. Grigg13Faculty of Medicine and Health Sciences, Universiti Malaysia SabahInfectious Diseases Society Kota Kinabalu - Menzies School of Health Research Clinical Research UnitInfectious Diseases Society Kota Kinabalu - Menzies School of Health Research Clinical Research UnitInfectious Diseases Society Kota Kinabalu - Menzies School of Health Research Clinical Research UnitInfectious Diseases Society Kota Kinabalu - Menzies School of Health Research Clinical Research UnitInfectious Diseases Society Kota Kinabalu - Menzies School of Health Research Clinical Research UnitMinistry of HealthState Public Health Laboratory, Sabah Department of HealthState Public Health Laboratory, Sabah Department of HealthInfectious Diseases Society Kota Kinabalu - Menzies School of Health Research Clinical Research UnitInfectious Diseases Society Kota Kinabalu - Menzies School of Health Research Clinical Research UnitInfectious Diseases Society Kota Kinabalu - Menzies School of Health Research Clinical Research UnitFaculty of Medicine and Health Sciences, Universiti Malaysia SabahInfectious Diseases Society Kota Kinabalu - Menzies School of Health Research Clinical Research UnitAbstract Background The monkey parasite Plasmodium knowlesi is an emerging public health issue in Southeast Asia. In Sabah, Malaysia, P. knowlesi is now the dominant cause of human malaria. Molecular detection methods for P. knowlesi are essential for accurate diagnosis and in monitoring progress towards malaria elimination of other Plasmodium species. However, recent commercially available PCR malaria kits have unpublished P. knowlesi gene targets or have not been evaluated against clinical samples. Methods Two real-time PCR methods currently used in Sabah for confirmatory malaria diagnosis and surveillance reporting were evaluated: the QuantiFast™ Multiplex PCR kit (Qiagen, Germany) targeting the P. knowlesi 18S SSU rRNA; and the abTES™ Malaria 5 qPCR II kit (AITbiotech, Singapore), with an undisclosed P. knowlesi gene target. Diagnostic accuracy was evaluated using 52 P. knowlesi, 25 Plasmodium vivax, 21 Plasmodium falciparum, and 10 Plasmodium malariae clinical isolates, and 26 malaria negative controls, and compared against a validated reference nested PCR assay. The limit of detection (LOD) for each PCR method and Plasmodium species was also evaluated. Results The sensitivity of the QuantiFast™ and abTES™ assays for detecting P. knowlesi was comparable at 98.1% (95% CI 89.7–100) and 100% (95% CI 93.2–100), respectively. Specificity of the QuantiFast™ and abTES™ for P. knowlesi was high at 98.8% (95% CI 93.4–100) for both assays. The QuantiFast™ assay demonstrated falsely-positive mixed Plasmodium species at low parasitaemias in both the primary and LOD analysis. Diagnostic accuracy of both PCR kits for detecting P. vivax, P. falciparum, and P. malariae was comparable to P. knowlesi. The abTES™ assay demonstrated a lower LOD for P. knowlesi of ≤ 0.125 parasites/µL compared to QuantiFast™ with a LOD of 20 parasites/µL. Hospital microscopy demonstrated a sensitivity of 78.8% (95% CI 65.3–88.9) and specificity of 80.4% (95% CI 67.6–89.8) compared to reference PCR for detecting P. knowlesi. Conclusion The QuantiFast™ and abTES™ commercial PCR kits performed well for the accurate detection of P. knowlesi infections. Although the QuantiFast™ kit is cheaper, the abTES™ kit demonstrated a lower LOD, supporting its use as a second-line referral-laboratory diagnostic tool in Sabah, Malaysia.http://link.springer.com/article/10.1186/s12936-020-03379-2Zoonotic malariaPlasmodium knowlesiReal-time polymerase chain reactionSabah Malaysia |
| spellingShingle | Nor Afizah Nuin Angelica F. Tan Yao Long Lew Kim A. Piera Timothy William Giri S. Rajahram Jenarun Jelip Jiloris F. Dony Rashidah Mohammad Daniel J. Cooper Bridget E. Barber Nicholas M. Anstey Tock H. Chua Matthew J. Grigg Comparative evaluation of two commercial real-time PCR kits (QuantiFast™ and abTES™) for the detection of Plasmodium knowlesi and other Plasmodium species in Sabah, Malaysia Malaria Journal Zoonotic malaria Plasmodium knowlesi Real-time polymerase chain reaction Sabah Malaysia |
| title | Comparative evaluation of two commercial real-time PCR kits (QuantiFast™ and abTES™) for the detection of Plasmodium knowlesi and other Plasmodium species in Sabah, Malaysia |
| title_full | Comparative evaluation of two commercial real-time PCR kits (QuantiFast™ and abTES™) for the detection of Plasmodium knowlesi and other Plasmodium species in Sabah, Malaysia |
| title_fullStr | Comparative evaluation of two commercial real-time PCR kits (QuantiFast™ and abTES™) for the detection of Plasmodium knowlesi and other Plasmodium species in Sabah, Malaysia |
| title_full_unstemmed | Comparative evaluation of two commercial real-time PCR kits (QuantiFast™ and abTES™) for the detection of Plasmodium knowlesi and other Plasmodium species in Sabah, Malaysia |
| title_short | Comparative evaluation of two commercial real-time PCR kits (QuantiFast™ and abTES™) for the detection of Plasmodium knowlesi and other Plasmodium species in Sabah, Malaysia |
| title_sort | comparative evaluation of two commercial real time pcr kits quantifast™ and abtes™ for the detection of plasmodium knowlesi and other plasmodium species in sabah malaysia |
| topic | Zoonotic malaria Plasmodium knowlesi Real-time polymerase chain reaction Sabah Malaysia |
| url | http://link.springer.com/article/10.1186/s12936-020-03379-2 |
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