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TaqMan real-time PCR for detection of pathogenic Leptospira spp. in canine clinical samples

Introduction: Canine leptospirosis has always been a differential diagnosis in dogs presenting with clinical signs and blood profiles associated with kidney and/or liver disease. The conventional polymerase chain reaction (PCR) provides diagnoses, but real-time PCR-based tests provide earlier conf...

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Main Authors: Abdul Rahman, Mohammad Sabri, Khor, Kuan Hua, Bejo, Siti Khairani, Lau, Seng Fong, Mazlan, Mazlina, Roslan, Mohd Azri, Md Ajat, Mohd Mokrish, Mohd Noor, Mohd Akmal
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Published: Sciendo 2023
Online Access:http://psasir.upm.edu.my/id/eprint/109212/
https://sciendo.com/article/10.2478/jvetres-2023-0024
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spelling my.upm.eprints.1092122024-08-27T04:06:49Z http://psasir.upm.edu.my/id/eprint/109212/ TaqMan real-time PCR for detection of pathogenic Leptospira spp. in canine clinical samples Abdul Rahman, Mohammad Sabri Khor, Kuan Hua Bejo, Siti Khairani Lau, Seng Fong Mazlan, Mazlina Roslan, Mohd Azri Md Ajat, Mohd Mokrish Mohd Noor, Mohd Akmal Introduction: Canine leptospirosis has always been a differential diagnosis in dogs presenting with clinical signs and blood profiles associated with kidney and/or liver disease. The conventional polymerase chain reaction (PCR) provides diagnoses, but real-time PCR-based tests provide earlier confirmation and determine the severity of infection, especially in the acute stage, allowing early detection for immediate treatment decisions. To our knowledge, real-time PCR has not been routinely adopted for clinical investigation in Malaysia. This study evaluated TaqMan real-time PCR (qPCR) assays diagnosing leptospirosis and compared their applicability to clinical samples from dogs with kidney and/or liver disease against a conventional PCR reference. Material and Methods: The qPCR assays were validated using existing leptospiral isolates. Whole blood and urine samples were analysed using a conventional PCR, LipL32(1) and LipL32(2) qPCRs and a microscopic agglutination test. The sensitivity and specificity of the qPCRs were determined. Results: The LipL32(1) qPCR assay had more diagnostic value than the LipL32(2) qPCR assay. Further evaluation of this assay revealed that it could detect as low as five DNA copies per reaction with high specificity for the tested leptospiral strains. No cross-amplification was observed with other organisms. Analysing the clinical samples, the LipL32(1) qPCR assay had 100.0% sensitivity and >75.0% specificity. Conclusion: The LipL32(1) qPCR assay is sensitive, specific and has the potential to be applied in future studies. Sciendo 2023-06 Article PeerReviewed Abdul Rahman, Mohammad Sabri and Khor, Kuan Hua and Bejo, Siti Khairani and Lau, Seng Fong and Mazlan, Mazlina and Roslan, Mohd Azri and Md Ajat, Mohd Mokrish and Mohd Noor, Mohd Akmal (2023) TaqMan real-time PCR for detection of pathogenic Leptospira spp. in canine clinical samples. Journal of Veterinary Research, 67 (2). pp. 187-195. ISSN 2450-8608 https://sciendo.com/article/10.2478/jvetres-2023-0024 10.2478/jvetres-2023-0024
institution Universiti Putra Malaysia
building UPM Library
collection Institutional Repository
continent Asia
country Malaysia
content_provider Universiti Putra Malaysia
content_source UPM Institutional Repository
url_provider http://psasir.upm.edu.my/
description Introduction: Canine leptospirosis has always been a differential diagnosis in dogs presenting with clinical signs and blood profiles associated with kidney and/or liver disease. The conventional polymerase chain reaction (PCR) provides diagnoses, but real-time PCR-based tests provide earlier confirmation and determine the severity of infection, especially in the acute stage, allowing early detection for immediate treatment decisions. To our knowledge, real-time PCR has not been routinely adopted for clinical investigation in Malaysia. This study evaluated TaqMan real-time PCR (qPCR) assays diagnosing leptospirosis and compared their applicability to clinical samples from dogs with kidney and/or liver disease against a conventional PCR reference. Material and Methods: The qPCR assays were validated using existing leptospiral isolates. Whole blood and urine samples were analysed using a conventional PCR, LipL32(1) and LipL32(2) qPCRs and a microscopic agglutination test. The sensitivity and specificity of the qPCRs were determined. Results: The LipL32(1) qPCR assay had more diagnostic value than the LipL32(2) qPCR assay. Further evaluation of this assay revealed that it could detect as low as five DNA copies per reaction with high specificity for the tested leptospiral strains. No cross-amplification was observed with other organisms. Analysing the clinical samples, the LipL32(1) qPCR assay had 100.0% sensitivity and >75.0% specificity. Conclusion: The LipL32(1) qPCR assay is sensitive, specific and has the potential to be applied in future studies.
format Article
author Abdul Rahman, Mohammad Sabri
Khor, Kuan Hua
Bejo, Siti Khairani
Lau, Seng Fong
Mazlan, Mazlina
Roslan, Mohd Azri
Md Ajat, Mohd Mokrish
Mohd Noor, Mohd Akmal
spellingShingle Abdul Rahman, Mohammad Sabri
Khor, Kuan Hua
Bejo, Siti Khairani
Lau, Seng Fong
Mazlan, Mazlina
Roslan, Mohd Azri
Md Ajat, Mohd Mokrish
Mohd Noor, Mohd Akmal
TaqMan real-time PCR for detection of pathogenic Leptospira spp. in canine clinical samples
author_facet Abdul Rahman, Mohammad Sabri
Khor, Kuan Hua
Bejo, Siti Khairani
Lau, Seng Fong
Mazlan, Mazlina
Roslan, Mohd Azri
Md Ajat, Mohd Mokrish
Mohd Noor, Mohd Akmal
author_sort Abdul Rahman, Mohammad Sabri
title TaqMan real-time PCR for detection of pathogenic Leptospira spp. in canine clinical samples
title_short TaqMan real-time PCR for detection of pathogenic Leptospira spp. in canine clinical samples
title_full TaqMan real-time PCR for detection of pathogenic Leptospira spp. in canine clinical samples
title_fullStr TaqMan real-time PCR for detection of pathogenic Leptospira spp. in canine clinical samples
title_full_unstemmed TaqMan real-time PCR for detection of pathogenic Leptospira spp. in canine clinical samples
title_sort taqman real-time pcr for detection of pathogenic leptospira spp. in canine clinical samples
publisher Sciendo
publishDate 2023
url http://psasir.upm.edu.my/id/eprint/109212/
https://sciendo.com/article/10.2478/jvetres-2023-0024
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score 13.211869

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