A comparison of two molecular methods for diagnosing leptospirosis from three different sample types in patients presenting with fever in Laos
<strong>Objectives</strong> To compare two molecular assays (rrs quantitative PCR (qPCR) versus a combined 16SrRNA and LipL32 qPCR) on different sample types for diagnosing leptospirosis in febrile patients presenting to Mahosot Hospital, Vientiane, Laos. <strong>Methods</stron...
| Main Authors: | , , , , , , , , , , , , , , , , , , |
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| Format: | Journal article |
| Language: | English |
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Elsevier
2017
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| _version_ | 1826288515545038848 |
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| author | Woods, K Nic-Fhogartaigh, C Arnold, C Boutthasavong, L Phuklia, W Lim, C Chanthongthip, A Tulsiani, S Craig, S Burns, M Weier, S Davong, V Sihalath, S Limmathurotsakul, D Dance, D Shetty, N Zambon, M Newton, P Dittrich, S |
| author_facet | Woods, K Nic-Fhogartaigh, C Arnold, C Boutthasavong, L Phuklia, W Lim, C Chanthongthip, A Tulsiani, S Craig, S Burns, M Weier, S Davong, V Sihalath, S Limmathurotsakul, D Dance, D Shetty, N Zambon, M Newton, P Dittrich, S |
| author_sort | Woods, K |
| collection | OXFORD |
| description | <strong>Objectives</strong> To compare two molecular assays (rrs quantitative PCR (qPCR) versus a combined 16SrRNA and LipL32 qPCR) on different sample types for diagnosing leptospirosis in febrile patients presenting to Mahosot Hospital, Vientiane, Laos. <strong>Methods</strong> Serum, buffy coat and urine samples were collected on admission, and follow-up serum ∼10 days later. Leptospira spp. culture and microscopic agglutination tests (MAT) were performed as reference standards. Bayesian latent class modelling was performed to estimate sensitivity and specificity of each diagnostic test. <strong>Results</strong> In all, 787 patients were included in the analysis: 4/787 (0.5%) were Leptospira culture positive, 30/787 (3.8%) were MAT positive, 76/787 (9.7%) were rrs qPCR positive and 20/787 (2.5%) were 16SrRNA/LipL32 qPCR positive for pathogenic Leptospira spp. in at least one sample. Estimated sensitivity and specificity (with 95% CI) of 16SrRNA/LipL32 qPCR on serum (53.9% (33.3%–81.8%); 99.6% (99.2%–100%)), buffy coat (58.8% (34.4%–90.9%); 99.9% (99.6%–100%)) and urine samples (45.0% (27.0%–66.7%); 99.6% (99.3%–100%)) were comparable with those of rrs qPCR, except specificity of 16SrRNA/LipL32 qPCR on urine samples was significantly higher (99.6% (99.3%–100%) vs. 92.5% (92.3%–92.8%), p <0.001). Sensitivities of MAT (16% (95% CI 6.3%–29.4%)) and culture (25% (95% CI 13.3%–44.4%)) were low. Mean positive Cq values showed that buffy coat samples were more frequently inhibitory to qPCR than either serum or urine (p <0.001). <strong>Conclusions</strong> Serum and urine are better samples for qPCR than buffy coat, and 16SrRNA/LipL32 qPCR performs better than rrs qPCR on urine. Quantitative PCR on admission is a reliable rapid diagnostic tool, performing better than MAT or culture, with significant implications for clinical and epidemiological investigations of this global neglected disease. |
| first_indexed | 2024-03-07T02:14:52Z |
| format | Journal article |
| id | oxford-uuid:a1dee7a7-7248-4c6a-aa14-06b823ef7f4a |
| institution | University of Oxford |
| language | English |
| last_indexed | 2024-03-07T02:14:52Z |
| publishDate | 2017 |
| publisher | Elsevier |
| record_format | dspace |
| spelling | oxford-uuid:a1dee7a7-7248-4c6a-aa14-06b823ef7f4a2022-03-27T02:16:23ZA comparison of two molecular methods for diagnosing leptospirosis from three different sample types in patients presenting with fever in LaosJournal articlehttp://purl.org/coar/resource_type/c_dcae04bcuuid:a1dee7a7-7248-4c6a-aa14-06b823ef7f4aEnglishSymplectic Elements at OxfordElsevier2017Woods, KNic-Fhogartaigh, CArnold, CBoutthasavong, LPhuklia, WLim, CChanthongthip, ATulsiani, SCraig, SBurns, MWeier, SDavong, VSihalath, SLimmathurotsakul, DDance, DShetty, NZambon, MNewton, PDittrich, S<strong>Objectives</strong> To compare two molecular assays (rrs quantitative PCR (qPCR) versus a combined 16SrRNA and LipL32 qPCR) on different sample types for diagnosing leptospirosis in febrile patients presenting to Mahosot Hospital, Vientiane, Laos. <strong>Methods</strong> Serum, buffy coat and urine samples were collected on admission, and follow-up serum ∼10 days later. Leptospira spp. culture and microscopic agglutination tests (MAT) were performed as reference standards. Bayesian latent class modelling was performed to estimate sensitivity and specificity of each diagnostic test. <strong>Results</strong> In all, 787 patients were included in the analysis: 4/787 (0.5%) were Leptospira culture positive, 30/787 (3.8%) were MAT positive, 76/787 (9.7%) were rrs qPCR positive and 20/787 (2.5%) were 16SrRNA/LipL32 qPCR positive for pathogenic Leptospira spp. in at least one sample. Estimated sensitivity and specificity (with 95% CI) of 16SrRNA/LipL32 qPCR on serum (53.9% (33.3%–81.8%); 99.6% (99.2%–100%)), buffy coat (58.8% (34.4%–90.9%); 99.9% (99.6%–100%)) and urine samples (45.0% (27.0%–66.7%); 99.6% (99.3%–100%)) were comparable with those of rrs qPCR, except specificity of 16SrRNA/LipL32 qPCR on urine samples was significantly higher (99.6% (99.3%–100%) vs. 92.5% (92.3%–92.8%), p <0.001). Sensitivities of MAT (16% (95% CI 6.3%–29.4%)) and culture (25% (95% CI 13.3%–44.4%)) were low. Mean positive Cq values showed that buffy coat samples were more frequently inhibitory to qPCR than either serum or urine (p <0.001). <strong>Conclusions</strong> Serum and urine are better samples for qPCR than buffy coat, and 16SrRNA/LipL32 qPCR performs better than rrs qPCR on urine. Quantitative PCR on admission is a reliable rapid diagnostic tool, performing better than MAT or culture, with significant implications for clinical and epidemiological investigations of this global neglected disease. |
| spellingShingle | Woods, K Nic-Fhogartaigh, C Arnold, C Boutthasavong, L Phuklia, W Lim, C Chanthongthip, A Tulsiani, S Craig, S Burns, M Weier, S Davong, V Sihalath, S Limmathurotsakul, D Dance, D Shetty, N Zambon, M Newton, P Dittrich, S A comparison of two molecular methods for diagnosing leptospirosis from three different sample types in patients presenting with fever in Laos |
| title | A comparison of two molecular methods for diagnosing leptospirosis from three different sample types in patients presenting with fever in Laos |
| title_full | A comparison of two molecular methods for diagnosing leptospirosis from three different sample types in patients presenting with fever in Laos |
| title_fullStr | A comparison of two molecular methods for diagnosing leptospirosis from three different sample types in patients presenting with fever in Laos |
| title_full_unstemmed | A comparison of two molecular methods for diagnosing leptospirosis from three different sample types in patients presenting with fever in Laos |
| title_short | A comparison of two molecular methods for diagnosing leptospirosis from three different sample types in patients presenting with fever in Laos |
| title_sort | comparison of two molecular methods for diagnosing leptospirosis from three different sample types in patients presenting with fever in laos |
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