A comparison of two molecular methods for diagnosing leptospirosis from three different sample types in patients presenting with fever in Laos

Kate Woods; Caoimhe Nic-Fhogartaigh; Catherine Arnold; Latzaniphone Boutthasavong; Weerawat Phuklia; Cherry Lim; Anisone Chanthongthip; Suhella M Tulsiani; Scott B Craig; Mary-Anne Burns; Steven L Weier; Viengmon Davong; Somsavanh Sihalath; Direk Limmathurotsakul; David A B Dance; Nandini Shetty; Maria Zambon; Paul N Newton; Sabine Dittrich

doi:10.1016/j.cmi.2017.10.017

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A comparison of two molecular methods for diagnosing leptospirosis from three different sample types in patients presenting with fever in Laos

Journal article

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Peer reviewed

A comparison of two molecular methods for diagnosing leptospirosis from three different sample types in patients presenting with fever in Laos

Kate Woods, Caoimhe Nic-Fhogartaigh, Catherine Arnold, Latzaniphone Boutthasavong, Weerawat Phuklia, Cherry Lim, Anisone Chanthongthip, Suhella M Tulsiani, Scott B Craig, Mary-Anne Burns, …

Clinical Microbiology and Infection, Vol.24(9), pp.1017.e1-1017.e7

2018

DOI: https://doi.org/10.1016/j.cmi.2017.10.017

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Abstract

Leptospirosis

Leptospira

molecular diagnosis

qPCR

Bayesian

Latent class model

serum

buffy coat

urine

Laos

Objectives: To compare two molecular assays (rrs qPCR vs. a combined 16SrRNA and LipL32 qPCR) on different sample types for diagnosing leptospirosis in febrile patients presenting to Mahosot Hospital, Vientiane, Laos. Methods: 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 modeling was performed to estimate sensitivity and specificity of each diagnostic test. Results: 787 patients were included in the analysis: 4/787 (0.5%) were Leptospira culture positive, 30/787 (3.8%) MAT positive, 76/787 (9.7%) rrs qPCR positive and 20/787 (2.5%) 16SrRNA/LipL32 qPCR positive for pathogenic Leptospira spp. in ≥1 sample. Estimated sensitivity and specificity (with 95% confidence intervals [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). Sensitivity 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). Conclusions: Serum and urine are better samples for qPCR than buffy coat, and 16SrRNA/LipL32 qPCR performs better than rrs qPCR on urine. qPCR 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.

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Title: A comparison of two molecular methods for diagnosing leptospirosis from three different sample types in patients presenting with fever in Laos
Authors: Kate Woods (Author) - Public Health England, United Kingdom
Caoimhe Nic-Fhogartaigh (Author) - Royal London Hospital, United Kingdom
Catherine Arnold (Author) - Public Health England, United Kingdom
Latzaniphone Boutthasavong (Author) - Mahosot Hospital, Vietnam
Weerawat Phuklia (Author) - Mahosot Hospital, Vietnam
Cherry Lim (Author) - Mahidol University, Thailand
Anisone Chanthongthip (Author) - Mahosot Hospital, Vietnam
Suhella M Tulsiani (Author) - Queensland Health Forensic and Scientific Service
Scott B Craig (Author) - University of the Sunshine Coast - Faculty of Science, Health, Education and Engineering
Mary-Anne Burns (Author) - Queensland Health Forensic and Scientific Service
Steven L Weier (Author) - Queensland University of Technology
Viengmon Davong (Author) - Mahosot University, Vietnam
Somsavanh Sihalath (Author) - Mahosot University, Vietnam
Direk Limmathurotsakul (Author) - Mahidol University, Thailand
David A B Dance (Author) - Mahosot University, Vietnam
Nandini Shetty (Author) - Public Health England, United Kingdom
Maria Zambon (Author) - Public Health England, United Kingdom
Paul N Newton (Author) - Mahosot University, Vietnam
Sabine Dittrich (Author) - Mahosot University, Vietnam
Publication details: Clinical Microbiology and Infection, Vol.24(9), pp.1017.e1-1017.e7
Publisher: Elsevier Ltd.
Date published: 2018
DOI: 10.1016/j.cmi.2017.10.017
ISSN: 1198-743X
Copyright note: Copyright © 2017 The Authors. Published by Elsevier Ltd on behalf of European Society of Clinical Microbiology and Infectious Diseases. This is an open access article underthe CC BY license (http://creativecommons.org/licenses/by/4.0/).Clinical Microbiology and Infection 24 (2018) 1017.e1e1017.e7
Organisation Unit: School of Science and Engineering - Legacy; University of the Sunshine Coast, Queensland
Language: English
Record Identifier: 99451172302621
Output Type: Journal article

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