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Development of multiplex PCR for neglected infectious diseases

Scrub typhus, murine typhus, and leptospirosis are widely neglected infectious diseases caused by Orientia tsutsugamushi, Rickettsia typhi, and pathogenic Leptospira spp., respectively. Patients usually present with non-specific symptoms and therefore are commonly diagnosed with acute undifferentiat...

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Autores principales: Sea-liang, Nutchanart, Sereemaspun, Amornpun, Patarakul, Kanitha, Gaywee, Jariyanart, Rodkvamtook, Wuttikon, Srisawat, Nattachai, Wacharaplusadee, Supaporn, Hemachudha, Thiravat
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6613674/
https://www.ncbi.nlm.nih.gov/pubmed/31283768
http://dx.doi.org/10.1371/journal.pntd.0007440
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author Sea-liang, Nutchanart
Sereemaspun, Amornpun
Patarakul, Kanitha
Gaywee, Jariyanart
Rodkvamtook, Wuttikon
Srisawat, Nattachai
Wacharaplusadee, Supaporn
Hemachudha, Thiravat
author_facet Sea-liang, Nutchanart
Sereemaspun, Amornpun
Patarakul, Kanitha
Gaywee, Jariyanart
Rodkvamtook, Wuttikon
Srisawat, Nattachai
Wacharaplusadee, Supaporn
Hemachudha, Thiravat
author_sort Sea-liang, Nutchanart
collection PubMed
description Scrub typhus, murine typhus, and leptospirosis are widely neglected infectious diseases caused by Orientia tsutsugamushi, Rickettsia typhi, and pathogenic Leptospira spp., respectively. Patients usually present with non-specific symptoms and therefore are commonly diagnosed with acute undifferentiated febrile illness. Consequently, patients face delayed treatment and increased mortality. Antibody-based serological test currently used as gold standard has limitations due to insufficient antibody titers, especially in the early phase of infection. In this study, we aimed to develop multiplex PCR to combine 3 primer pairs that target specific genes encoding 56-kDa TSA of O. tsutsugamushi, 17-kDa antigen of R. typhi, and LipL32 of L. Interrogans and evaluate its performance in comparison to the standard serological tests. Using EDTA blood samples of known patients, the sensitivity and specificity of our multiplex PCR was 100% and 70%, respectively. In addition, the assay was able to diagnose the co-infection of scrub typhus and leptospirosis. The assay may be useful in identifying causative agents during the early phase of these diseases, enabling prompt and appropriate treatment.
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spelling pubmed-66136742019-07-23 Development of multiplex PCR for neglected infectious diseases Sea-liang, Nutchanart Sereemaspun, Amornpun Patarakul, Kanitha Gaywee, Jariyanart Rodkvamtook, Wuttikon Srisawat, Nattachai Wacharaplusadee, Supaporn Hemachudha, Thiravat PLoS Negl Trop Dis Research Article Scrub typhus, murine typhus, and leptospirosis are widely neglected infectious diseases caused by Orientia tsutsugamushi, Rickettsia typhi, and pathogenic Leptospira spp., respectively. Patients usually present with non-specific symptoms and therefore are commonly diagnosed with acute undifferentiated febrile illness. Consequently, patients face delayed treatment and increased mortality. Antibody-based serological test currently used as gold standard has limitations due to insufficient antibody titers, especially in the early phase of infection. In this study, we aimed to develop multiplex PCR to combine 3 primer pairs that target specific genes encoding 56-kDa TSA of O. tsutsugamushi, 17-kDa antigen of R. typhi, and LipL32 of L. Interrogans and evaluate its performance in comparison to the standard serological tests. Using EDTA blood samples of known patients, the sensitivity and specificity of our multiplex PCR was 100% and 70%, respectively. In addition, the assay was able to diagnose the co-infection of scrub typhus and leptospirosis. The assay may be useful in identifying causative agents during the early phase of these diseases, enabling prompt and appropriate treatment. Public Library of Science 2019-07-08 /pmc/articles/PMC6613674/ /pubmed/31283768 http://dx.doi.org/10.1371/journal.pntd.0007440 Text en © 2019 Sea-liang et al http://creativecommons.org/licenses/by/4.0/ This is an open access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
spellingShingle Research Article
Sea-liang, Nutchanart
Sereemaspun, Amornpun
Patarakul, Kanitha
Gaywee, Jariyanart
Rodkvamtook, Wuttikon
Srisawat, Nattachai
Wacharaplusadee, Supaporn
Hemachudha, Thiravat
Development of multiplex PCR for neglected infectious diseases
title Development of multiplex PCR for neglected infectious diseases
title_full Development of multiplex PCR for neglected infectious diseases
title_fullStr Development of multiplex PCR for neglected infectious diseases
title_full_unstemmed Development of multiplex PCR for neglected infectious diseases
title_short Development of multiplex PCR for neglected infectious diseases
title_sort development of multiplex pcr for neglected infectious diseases
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6613674/
https://www.ncbi.nlm.nih.gov/pubmed/31283768
http://dx.doi.org/10.1371/journal.pntd.0007440
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