Rapid detection of pathogenic leptospires by lyophilized reagent-based Polymerase Chain Reaction.

A simple and reliable tool for the early diagnosis of leptospirosis is urgently needed. We report the development of a lyophilized reagent-based polymerase chain reaction (PCR) assay targeting lipL32 gene, which is present only in pathogenic leptospires. To determine the effectiveness of the newly d...

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Main Authors: Lee, S.V., Tai, E.S., Mutalib, Abdul Rahim, Bejo, Siti Khairani, Bahaman, Abdul Rani
Format: Article
Language:English
English
Published: Malaysian Society of Parasitology and Tropical Medicine 2011
Online Access:http://psasir.upm.edu.my/id/eprint/25403/
http://psasir.upm.edu.my/id/eprint/25403/1/Rapid%20detection%20of%20pathogenic%20leptospires%20by%20lyophilized%20reagent.pdf
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author Lee, S.V.
Tai, E.S.
Mutalib, Abdul Rahim
Bejo, Siti Khairani
Bahaman, Abdul Rani
author_facet Lee, S.V.
Tai, E.S.
Mutalib, Abdul Rahim
Bejo, Siti Khairani
Bahaman, Abdul Rani
author_sort Lee, S.V.
building UPM Institutional Repository
collection Online Access
description A simple and reliable tool for the early diagnosis of leptospirosis is urgently needed. We report the development of a lyophilized reagent-based polymerase chain reaction (PCR) assay targeting lipL32 gene, which is present only in pathogenic leptospires. To determine the effectiveness of the newly developed assay in the early diagnosis of leptospirosis, the sensitivity and specificity was evaluated. In simulated clinical samples, the assay was able to detect 10 2 and 10 3 leptospires/ml in spiked urine and blood samples, respectively. In experimentally infected animals, leptospiral DNA could be detected in blood and lung samples as early as Day 1 post infection. This assay was also shown to be stable and remained sensitive for up to five months at ambient temperature. Hence, this lyophilized reagent-based PCR assay with high specificity, sensitivity and stability would provide a simple, rapid and reliable method in diagnosing acute leptospirosis, especially in the field of veterinary medicine.
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spelling upm-254032015-10-02T00:33:59Z http://psasir.upm.edu.my/id/eprint/25403/ Rapid detection of pathogenic leptospires by lyophilized reagent-based Polymerase Chain Reaction. Lee, S.V. Tai, E.S. Mutalib, Abdul Rahim Bejo, Siti Khairani Bahaman, Abdul Rani A simple and reliable tool for the early diagnosis of leptospirosis is urgently needed. We report the development of a lyophilized reagent-based polymerase chain reaction (PCR) assay targeting lipL32 gene, which is present only in pathogenic leptospires. To determine the effectiveness of the newly developed assay in the early diagnosis of leptospirosis, the sensitivity and specificity was evaluated. In simulated clinical samples, the assay was able to detect 10 2 and 10 3 leptospires/ml in spiked urine and blood samples, respectively. In experimentally infected animals, leptospiral DNA could be detected in blood and lung samples as early as Day 1 post infection. This assay was also shown to be stable and remained sensitive for up to five months at ambient temperature. Hence, this lyophilized reagent-based PCR assay with high specificity, sensitivity and stability would provide a simple, rapid and reliable method in diagnosing acute leptospirosis, especially in the field of veterinary medicine. Malaysian Society of Parasitology and Tropical Medicine 2011-12 Article PeerReviewed application/pdf en http://psasir.upm.edu.my/id/eprint/25403/1/Rapid%20detection%20of%20pathogenic%20leptospires%20by%20lyophilized%20reagent.pdf Lee, S.V. and Tai, E.S. and Mutalib, Abdul Rahim and Bejo, Siti Khairani and Bahaman, Abdul Rani (2011) Rapid detection of pathogenic leptospires by lyophilized reagent-based Polymerase Chain Reaction. Tropical Biomedicine, 28 (3). pp. 497-505. ISSN 0127-5720 English
spellingShingle Lee, S.V.
Tai, E.S.
Mutalib, Abdul Rahim
Bejo, Siti Khairani
Bahaman, Abdul Rani
Rapid detection of pathogenic leptospires by lyophilized reagent-based Polymerase Chain Reaction.
title Rapid detection of pathogenic leptospires by lyophilized reagent-based Polymerase Chain Reaction.
title_full Rapid detection of pathogenic leptospires by lyophilized reagent-based Polymerase Chain Reaction.
title_fullStr Rapid detection of pathogenic leptospires by lyophilized reagent-based Polymerase Chain Reaction.
title_full_unstemmed Rapid detection of pathogenic leptospires by lyophilized reagent-based Polymerase Chain Reaction.
title_short Rapid detection of pathogenic leptospires by lyophilized reagent-based Polymerase Chain Reaction.
title_sort rapid detection of pathogenic leptospires by lyophilized reagent-based polymerase chain reaction.
url http://psasir.upm.edu.my/id/eprint/25403/
http://psasir.upm.edu.my/id/eprint/25403/1/Rapid%20detection%20of%20pathogenic%20leptospires%20by%20lyophilized%20reagent.pdf