Rapid Diagnostic Tests for the Detection of the Four Dengue Virus Serotypes in Clinically Relevant Matrices
ABSTRACT The efficient and accurate diagnosis of dengue, a major mosquito-borne disease, is of primary importance for clinical care, surveillance, and outbreak control. The identification of specific dengue virus serotype 1 (DENV-1) to DENV-4 can help in understanding the transmission dynamics and s...
| Main Authors: | , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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American Society for Microbiology
2023-02-01
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| Series: | Microbiology Spectrum |
| Subjects: | |
| Online Access: | https://journals.asm.org/doi/10.1128/spectrum.02796-22 |
| _version_ | 1811164368810803200 |
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| author | Nina M. Pollak Malin Olsson Madeeha Ahmed Javier Tan George Lim Yin Xiang Setoh Judith Chui Ching Wong Yee Ling Lai Jody Hobson-Peters Joanne Macdonald David McMillan |
| author_facet | Nina M. Pollak Malin Olsson Madeeha Ahmed Javier Tan George Lim Yin Xiang Setoh Judith Chui Ching Wong Yee Ling Lai Jody Hobson-Peters Joanne Macdonald David McMillan |
| author_sort | Nina M. Pollak |
| collection | DOAJ |
| description | ABSTRACT The efficient and accurate diagnosis of dengue, a major mosquito-borne disease, is of primary importance for clinical care, surveillance, and outbreak control. The identification of specific dengue virus serotype 1 (DENV-1) to DENV-4 can help in understanding the transmission dynamics and spread of dengue disease. The four rapid low-resource serotype-specific dengue tests use a simple sample preparation reagent followed by reverse transcription-isothermal recombinase polymerase amplification (RT-RPA) combined with lateral flow detection (LFD) technology. Results are obtained directly from clinical sample matrices in 35 min, requiring only a heating block and pipettes for liquid handling. In addition, we demonstrate that the rapid sample preparation step inactivates DENV, improving laboratory safety. Human plasma and serum were spiked with DENV, and DENV was detected with analytical sensitivities of 333 to 22,500 median tissue culture infectious doses (TCID50)/mL. The analytical sensitivities in blood were 94,000 to 333,000 TCID50/mL. Analytical specificity testing confirmed that each test could detect multiple serotype-specific strains but did not respond to strains of other serotypes, closely related flaviviruses, or chikungunya virus. Clinical testing on 80 human serum samples demonstrated test specificities of between 94 and 100%, with a DENV-2 test sensitivity of 100%, detecting down to 0.004 PFU/μL, similar to the sensitivity of the PCR test; the other DENV tests detected down to 0.03 to 10.9 PFU/μL. Collectively, our data suggest that some of our rapid dengue serotyping tests provide a potential alternative to conventional labor-intensive RT-quantitative PCR (RT-qPCR) detection, which requires expensive thermal cycling instrumentation, technical expertise, and prolonged testing times. Our tests provide performance and speed without compromising specificity in human plasma and serum and could become promising tools for the detection of high DENV loads in resource-limited settings. IMPORTANCE The efficient and accurate diagnosis of dengue, a major mosquito-borne disease, is of primary importance for clinical care, surveillance, and outbreak control. This study describes the evaluation of four rapid low-resource serotype-specific dengue tests for the detection of specific DENV serotypes in clinical sample matrices. The tests use a simple sample preparation reagent followed by reverse transcription-isothermal recombinase polymerase amplification (RT-RPA) combined with lateral flow detection (LFD) technology. These tests have several advantages compared to RT-qPCR detection, such as a simple workflow, rapid sample processing and turnaround times (35 min from sample preparation to detection), minimal equipment needs, and improved laboratory safety through the inactivation of the virus during the sample preparation step. The low-resource formats of these rapid dengue serotyping tests have the potential to support effective dengue disease surveillance and enhance the diagnostic testing capacity in resource-limited countries with both endemic dengue and intense coronavirus disease 2019 (COVID-19) transmission. |
| first_indexed | 2024-04-10T15:21:30Z |
| format | Article |
| id | doaj.art-5ea13e94551a4c309371a8d91d467c51 |
| institution | Directory Open Access Journal |
| issn | 2165-0497 |
| language | English |
| last_indexed | 2024-04-10T15:21:30Z |
| publishDate | 2023-02-01 |
| publisher | American Society for Microbiology |
| record_format | Article |
| series | Microbiology Spectrum |
| spelling | doaj.art-5ea13e94551a4c309371a8d91d467c512023-02-14T14:15:49ZengAmerican Society for MicrobiologyMicrobiology Spectrum2165-04972023-02-0111110.1128/spectrum.02796-22Rapid Diagnostic Tests for the Detection of the Four Dengue Virus Serotypes in Clinically Relevant MatricesNina M. Pollak0Malin Olsson1Madeeha Ahmed2Javier Tan3George Lim4Yin Xiang Setoh5Judith Chui Ching Wong6Yee Ling Lai7Jody Hobson-Peters8Joanne Macdonald9David McMillan10Centre for Bioinnovation, University of the Sunshine Coast, Sippy Downs, Queensland, AustraliaCentre for Bioinnovation, University of the Sunshine Coast, Sippy Downs, Queensland, AustraliaCentre for Bioinnovation, University of the Sunshine Coast, Sippy Downs, Queensland, AustraliaEnvironmental Health Institute, National Environment Agency, Singapore, SingaporeEnvironmental Health Institute, National Environment Agency, Singapore, SingaporeEnvironmental Health Institute, National Environment Agency, Singapore, SingaporeEnvironmental Health Institute, National Environment Agency, Singapore, SingaporeEnvironmental Health Institute, National Environment Agency, Singapore, SingaporeSchool of Chemistry and Molecular Biosciences, The University of Queensland, St. Lucia, Queensland, AustraliaDMTC Ltd., Kew, Victoria, AustraliaCentre for Bioinnovation, University of the Sunshine Coast, Sippy Downs, Queensland, AustraliaABSTRACT The efficient and accurate diagnosis of dengue, a major mosquito-borne disease, is of primary importance for clinical care, surveillance, and outbreak control. The identification of specific dengue virus serotype 1 (DENV-1) to DENV-4 can help in understanding the transmission dynamics and spread of dengue disease. The four rapid low-resource serotype-specific dengue tests use a simple sample preparation reagent followed by reverse transcription-isothermal recombinase polymerase amplification (RT-RPA) combined with lateral flow detection (LFD) technology. Results are obtained directly from clinical sample matrices in 35 min, requiring only a heating block and pipettes for liquid handling. In addition, we demonstrate that the rapid sample preparation step inactivates DENV, improving laboratory safety. Human plasma and serum were spiked with DENV, and DENV was detected with analytical sensitivities of 333 to 22,500 median tissue culture infectious doses (TCID50)/mL. The analytical sensitivities in blood were 94,000 to 333,000 TCID50/mL. Analytical specificity testing confirmed that each test could detect multiple serotype-specific strains but did not respond to strains of other serotypes, closely related flaviviruses, or chikungunya virus. Clinical testing on 80 human serum samples demonstrated test specificities of between 94 and 100%, with a DENV-2 test sensitivity of 100%, detecting down to 0.004 PFU/μL, similar to the sensitivity of the PCR test; the other DENV tests detected down to 0.03 to 10.9 PFU/μL. Collectively, our data suggest that some of our rapid dengue serotyping tests provide a potential alternative to conventional labor-intensive RT-quantitative PCR (RT-qPCR) detection, which requires expensive thermal cycling instrumentation, technical expertise, and prolonged testing times. Our tests provide performance and speed without compromising specificity in human plasma and serum and could become promising tools for the detection of high DENV loads in resource-limited settings. IMPORTANCE The efficient and accurate diagnosis of dengue, a major mosquito-borne disease, is of primary importance for clinical care, surveillance, and outbreak control. This study describes the evaluation of four rapid low-resource serotype-specific dengue tests for the detection of specific DENV serotypes in clinical sample matrices. The tests use a simple sample preparation reagent followed by reverse transcription-isothermal recombinase polymerase amplification (RT-RPA) combined with lateral flow detection (LFD) technology. These tests have several advantages compared to RT-qPCR detection, such as a simple workflow, rapid sample processing and turnaround times (35 min from sample preparation to detection), minimal equipment needs, and improved laboratory safety through the inactivation of the virus during the sample preparation step. The low-resource formats of these rapid dengue serotyping tests have the potential to support effective dengue disease surveillance and enhance the diagnostic testing capacity in resource-limited countries with both endemic dengue and intense coronavirus disease 2019 (COVID-19) transmission.https://journals.asm.org/doi/10.1128/spectrum.02796-22dengue virusNS5 generapid sample preparationrecombinase polymerase amplificationisothermal amplificationlateral flow detection |
| spellingShingle | Nina M. Pollak Malin Olsson Madeeha Ahmed Javier Tan George Lim Yin Xiang Setoh Judith Chui Ching Wong Yee Ling Lai Jody Hobson-Peters Joanne Macdonald David McMillan Rapid Diagnostic Tests for the Detection of the Four Dengue Virus Serotypes in Clinically Relevant Matrices Microbiology Spectrum dengue virus NS5 gene rapid sample preparation recombinase polymerase amplification isothermal amplification lateral flow detection |
| title | Rapid Diagnostic Tests for the Detection of the Four Dengue Virus Serotypes in Clinically Relevant Matrices |
| title_full | Rapid Diagnostic Tests for the Detection of the Four Dengue Virus Serotypes in Clinically Relevant Matrices |
| title_fullStr | Rapid Diagnostic Tests for the Detection of the Four Dengue Virus Serotypes in Clinically Relevant Matrices |
| title_full_unstemmed | Rapid Diagnostic Tests for the Detection of the Four Dengue Virus Serotypes in Clinically Relevant Matrices |
| title_short | Rapid Diagnostic Tests for the Detection of the Four Dengue Virus Serotypes in Clinically Relevant Matrices |
| title_sort | rapid diagnostic tests for the detection of the four dengue virus serotypes in clinically relevant matrices |
| topic | dengue virus NS5 gene rapid sample preparation recombinase polymerase amplification isothermal amplification lateral flow detection |
| url | https://journals.asm.org/doi/10.1128/spectrum.02796-22 |
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