Development and Evaluation of a Nested PCR for Improved Diagnosis and Genetic Analysis of Peste des Petits Ruminants Virus (PPRV) for Future Use in Nascent PPR Eradication Programme

Peste des petits ruminants (PPR) is a highly contagious viral disease of small ruminants caused by PPR virus (PPRV). PPR is endemic in Asia, the Middle East and across large areas of Africa and is currently targeted for global eradication by 2030. The virus exists as four different lineages that are...

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Autores principales:	Mana Mahapatra, Martin Mayora Neto, Asha Khunti, Felix Njeumi, Satya Parida
Formato:	article
Lenguaje:	EN
Publicado:	MDPI AG 2021
Materias:	peste des petits ruminants virus PPR RT-PCR rapid detection diagnostic genetic analysis Veterinary medicine SF600-1100 Zoology QL1-991
Acceso en línea:	https://doaj.org/article/206d1a53cf1f4104a1488473ff4bd88c
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spelling	oai:doaj.org-article:206d1a53cf1f4104a1488473ff4bd88c2021-11-25T16:17:58ZDevelopment and Evaluation of a Nested PCR for Improved Diagnosis and Genetic Analysis of Peste des Petits Ruminants Virus (PPRV) for Future Use in Nascent PPR Eradication Programme10.3390/ani111131702076-2615https://doaj.org/article/206d1a53cf1f4104a1488473ff4bd88c2021-11-01T00:00:00Zhttps://www.mdpi.com/2076-2615/11/11/3170https://doaj.org/toc/2076-2615Peste des petits ruminants (PPR) is a highly contagious viral disease of small ruminants caused by PPR virus (PPRV). PPR is endemic in Asia, the Middle East and across large areas of Africa and is currently targeted for global eradication by 2030. The virus exists as four different lineages that are usually limited to specific geographical areas. However, recent reports of spread of PPRV, in particular of lineage IV viruses to infection-free countries and previously PPR endemic areas are noteworthy. A rapid and accurate laboratory diagnosis and reports on its epidemiological linkage for virus spread play a major role in the effective control and eradication of the disease. Currently, molecular assays, including conventional reverse transcription-polymerase chain reaction (RT-PCR) and real-time RT-PCR (RT-qPCR) are usually used for diagnosis of PPR while the sequencing of part of the nucleocapsid gene is usually carried out for the viral lineage identification. However, it is difficult to diagnose and sequence the genetic material if the animal excreted a low level of virus at the initial stage of infection or if the PPRV is degraded during the long-distance transportation of samples to the reference laboratories. This study describes the development of a novel nested RT-PCR assay for the detection of the PPRV nucleic acid by targeting the N-protein gene, compares the performance of the assay with the existing conventional RT-PCR and also provides good-quality DNA suitable for sequencing in order to identify circulating lineages. The assay was evaluated using cell culture propagated PPRVs, field samples from clinically infected animals and samples from experimentally infected animals encompassing all four lineages (I–IV) of PPRV. This assay provides a solution with an easy, accurate, rapid and cost-effective PPR diagnostic and partial genome sequencing for use in resource-limited settings.Mana MahapatraMartin Mayora NetoAsha KhuntiFelix NjeumiSatya ParidaMDPI AGarticlepeste des petits ruminants virusPPRRT-PCRrapid detectiondiagnosticgenetic analysisVeterinary medicineSF600-1100ZoologyQL1-991ENAnimals, Vol 11, Iss 3170, p 3170 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	peste des petits ruminants virus PPR RT-PCR rapid detection diagnostic genetic analysis Veterinary medicine SF600-1100 Zoology QL1-991
spellingShingle	peste des petits ruminants virus PPR RT-PCR rapid detection diagnostic genetic analysis Veterinary medicine SF600-1100 Zoology QL1-991 Mana Mahapatra Martin Mayora Neto Asha Khunti Felix Njeumi Satya Parida Development and Evaluation of a Nested PCR for Improved Diagnosis and Genetic Analysis of Peste des Petits Ruminants Virus (PPRV) for Future Use in Nascent PPR Eradication Programme
description	Peste des petits ruminants (PPR) is a highly contagious viral disease of small ruminants caused by PPR virus (PPRV). PPR is endemic in Asia, the Middle East and across large areas of Africa and is currently targeted for global eradication by 2030. The virus exists as four different lineages that are usually limited to specific geographical areas. However, recent reports of spread of PPRV, in particular of lineage IV viruses to infection-free countries and previously PPR endemic areas are noteworthy. A rapid and accurate laboratory diagnosis and reports on its epidemiological linkage for virus spread play a major role in the effective control and eradication of the disease. Currently, molecular assays, including conventional reverse transcription-polymerase chain reaction (RT-PCR) and real-time RT-PCR (RT-qPCR) are usually used for diagnosis of PPR while the sequencing of part of the nucleocapsid gene is usually carried out for the viral lineage identification. However, it is difficult to diagnose and sequence the genetic material if the animal excreted a low level of virus at the initial stage of infection or if the PPRV is degraded during the long-distance transportation of samples to the reference laboratories. This study describes the development of a novel nested RT-PCR assay for the detection of the PPRV nucleic acid by targeting the N-protein gene, compares the performance of the assay with the existing conventional RT-PCR and also provides good-quality DNA suitable for sequencing in order to identify circulating lineages. The assay was evaluated using cell culture propagated PPRVs, field samples from clinically infected animals and samples from experimentally infected animals encompassing all four lineages (I–IV) of PPRV. This assay provides a solution with an easy, accurate, rapid and cost-effective PPR diagnostic and partial genome sequencing for use in resource-limited settings.
format	article
author	Mana Mahapatra Martin Mayora Neto Asha Khunti Felix Njeumi Satya Parida
author_facet	Mana Mahapatra Martin Mayora Neto Asha Khunti Felix Njeumi Satya Parida
author_sort	Mana Mahapatra
title	Development and Evaluation of a Nested PCR for Improved Diagnosis and Genetic Analysis of Peste des Petits Ruminants Virus (PPRV) for Future Use in Nascent PPR Eradication Programme
title_short	Development and Evaluation of a Nested PCR for Improved Diagnosis and Genetic Analysis of Peste des Petits Ruminants Virus (PPRV) for Future Use in Nascent PPR Eradication Programme
title_full	Development and Evaluation of a Nested PCR for Improved Diagnosis and Genetic Analysis of Peste des Petits Ruminants Virus (PPRV) for Future Use in Nascent PPR Eradication Programme
title_fullStr	Development and Evaluation of a Nested PCR for Improved Diagnosis and Genetic Analysis of Peste des Petits Ruminants Virus (PPRV) for Future Use in Nascent PPR Eradication Programme
title_full_unstemmed	Development and Evaluation of a Nested PCR for Improved Diagnosis and Genetic Analysis of Peste des Petits Ruminants Virus (PPRV) for Future Use in Nascent PPR Eradication Programme
title_sort	development and evaluation of a nested pcr for improved diagnosis and genetic analysis of peste des petits ruminants virus (pprv) for future use in nascent ppr eradication programme
publisher	MDPI AG
publishDate	2021
url	https://doaj.org/article/206d1a53cf1f4104a1488473ff4bd88c
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Development and Evaluation of a Nested PCR for Improved Diagnosis and Genetic Analysis of Peste des Petits Ruminants Virus (PPRV) for Future Use in Nascent PPR Eradication Programme

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