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Comparison of colorimetric loop-mediated isothermal amplification kit and reverse transcription-polymerase chain reaction in the diagnosis of peste des petits ruminants in sheep and goats in Southeast Nigeria

BACKGROUND AND AIM: Peste des petits ruminants (PPR) is an acute, extremely contagious transboundary viral disease of small ruminants with severe economic consequences, caused by PPR virus. Cost-effective and rapid diagnosis of the disease is essential for prompt management and control. This study a...

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Autores principales: Chukwudi, Ijeoma Chekwube, Ogbu, Kenneth Ikejiofor, Luka, Pam Dachung, Malesa, Refiloe Petunia, Heath, Livio Edward, Ugochukwu, Emmanuel Ikenna, Chah, Kennedy Foinkfu
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Veterinary World 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7750212/
https://www.ncbi.nlm.nih.gov/pubmed/33363327
http://dx.doi.org/10.14202/vetworld.2020.2358-2363
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author Chukwudi, Ijeoma Chekwube
Ogbu, Kenneth Ikejiofor
Luka, Pam Dachung
Malesa, Refiloe Petunia
Heath, Livio Edward
Ugochukwu, Emmanuel Ikenna
Chah, Kennedy Foinkfu
author_facet Chukwudi, Ijeoma Chekwube
Ogbu, Kenneth Ikejiofor
Luka, Pam Dachung
Malesa, Refiloe Petunia
Heath, Livio Edward
Ugochukwu, Emmanuel Ikenna
Chah, Kennedy Foinkfu
author_sort Chukwudi, Ijeoma Chekwube
collection PubMed
description BACKGROUND AND AIM: Peste des petits ruminants (PPR) is an acute, extremely contagious transboundary viral disease of small ruminants with severe economic consequences, caused by PPR virus. Cost-effective and rapid diagnosis of the disease is essential for prompt management and control. This study aimed to compare the application of a commercial colorimetric loop-mediated isothermal amplification (cLAMP) kit and reverse transcriptase-polymerase chain reaction (RT-PCR) in the diagnosis of PPR in sheep and goats in Southeast Nigeria. MATERIALS AND METHODS: Nasal swab samples were collected from West African Dwarf sheep and goats showing clinical signs suggestive of PPR (n=80) and those without any clinical signs (n=140) of the disease. The diagnosis was achieved through detection of PPR viral genome in the samples using a cLAMP kit and RT-PCR. cLAMP assay was done directly on nasal swab samples without ribosomal nucleic acid extraction. A set of six primers targeting the matrix gene protein was used for the cLAMP assay. RESULTS: PPR viral genome was detected by both cLAMP and RT-PCR in 51 (63.8%) of the 80 samples from sheep and goats with signs suggestive of PPR while 14 (10%) of those without signs tested positive for PPR by both assay methods. There was a 100% agreement in the cLAMP and RT-PCR results. However, cLAMP was a faster, easier, and less expensive method compared to RT-PCR. CONCLUSION: The cLAMP assay demonstrates the potential for a point of care diagnosis in the field and a valuable diagnostic tool in areas with poor electricity supply as well as in a less equipped diagnostic laboratory. Since the reagents are affordable, cLAMP can be a diagnostic tool of choice in the detection and surveillance of PPR virus in countries with limited resources.
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spelling pubmed-77502122020-12-23 Comparison of colorimetric loop-mediated isothermal amplification kit and reverse transcription-polymerase chain reaction in the diagnosis of peste des petits ruminants in sheep and goats in Southeast Nigeria Chukwudi, Ijeoma Chekwube Ogbu, Kenneth Ikejiofor Luka, Pam Dachung Malesa, Refiloe Petunia Heath, Livio Edward Ugochukwu, Emmanuel Ikenna Chah, Kennedy Foinkfu Vet World Research Article BACKGROUND AND AIM: Peste des petits ruminants (PPR) is an acute, extremely contagious transboundary viral disease of small ruminants with severe economic consequences, caused by PPR virus. Cost-effective and rapid diagnosis of the disease is essential for prompt management and control. This study aimed to compare the application of a commercial colorimetric loop-mediated isothermal amplification (cLAMP) kit and reverse transcriptase-polymerase chain reaction (RT-PCR) in the diagnosis of PPR in sheep and goats in Southeast Nigeria. MATERIALS AND METHODS: Nasal swab samples were collected from West African Dwarf sheep and goats showing clinical signs suggestive of PPR (n=80) and those without any clinical signs (n=140) of the disease. The diagnosis was achieved through detection of PPR viral genome in the samples using a cLAMP kit and RT-PCR. cLAMP assay was done directly on nasal swab samples without ribosomal nucleic acid extraction. A set of six primers targeting the matrix gene protein was used for the cLAMP assay. RESULTS: PPR viral genome was detected by both cLAMP and RT-PCR in 51 (63.8%) of the 80 samples from sheep and goats with signs suggestive of PPR while 14 (10%) of those without signs tested positive for PPR by both assay methods. There was a 100% agreement in the cLAMP and RT-PCR results. However, cLAMP was a faster, easier, and less expensive method compared to RT-PCR. CONCLUSION: The cLAMP assay demonstrates the potential for a point of care diagnosis in the field and a valuable diagnostic tool in areas with poor electricity supply as well as in a less equipped diagnostic laboratory. Since the reagents are affordable, cLAMP can be a diagnostic tool of choice in the detection and surveillance of PPR virus in countries with limited resources. Veterinary World 2020-11 2020-11-07 /pmc/articles/PMC7750212/ /pubmed/33363327 http://dx.doi.org/10.14202/vetworld.2020.2358-2363 Text en Copyright: © Chukwudi, et al. http://creativecommons.org/licenses/by/4.0 Open Access. This article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.
spellingShingle Research Article
Chukwudi, Ijeoma Chekwube
Ogbu, Kenneth Ikejiofor
Luka, Pam Dachung
Malesa, Refiloe Petunia
Heath, Livio Edward
Ugochukwu, Emmanuel Ikenna
Chah, Kennedy Foinkfu
Comparison of colorimetric loop-mediated isothermal amplification kit and reverse transcription-polymerase chain reaction in the diagnosis of peste des petits ruminants in sheep and goats in Southeast Nigeria
title Comparison of colorimetric loop-mediated isothermal amplification kit and reverse transcription-polymerase chain reaction in the diagnosis of peste des petits ruminants in sheep and goats in Southeast Nigeria
title_full Comparison of colorimetric loop-mediated isothermal amplification kit and reverse transcription-polymerase chain reaction in the diagnosis of peste des petits ruminants in sheep and goats in Southeast Nigeria
title_fullStr Comparison of colorimetric loop-mediated isothermal amplification kit and reverse transcription-polymerase chain reaction in the diagnosis of peste des petits ruminants in sheep and goats in Southeast Nigeria
title_full_unstemmed Comparison of colorimetric loop-mediated isothermal amplification kit and reverse transcription-polymerase chain reaction in the diagnosis of peste des petits ruminants in sheep and goats in Southeast Nigeria
title_short Comparison of colorimetric loop-mediated isothermal amplification kit and reverse transcription-polymerase chain reaction in the diagnosis of peste des petits ruminants in sheep and goats in Southeast Nigeria
title_sort comparison of colorimetric loop-mediated isothermal amplification kit and reverse transcription-polymerase chain reaction in the diagnosis of peste des petits ruminants in sheep and goats in southeast nigeria
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7750212/
https://www.ncbi.nlm.nih.gov/pubmed/33363327
http://dx.doi.org/10.14202/vetworld.2020.2358-2363
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