Recent advances in the laboratory diagnosis of peste des petits ruminants

2021 ◽  
Vol 77 (05) ◽  
pp. 226-231
Author(s):  
WIESŁAW NIEDBALSKI ◽  
ANDRZEJ FITZNER ◽  
KRZYSZTOF BULENGER ◽  
ANDRZEJ KĘSY
Keyword(s):  
Reverse Transcription ◽  
Animal Health ◽  
Clinical Signs ◽  
Small Ruminants ◽  
Clinical Diagnostics ◽  
Molecular Diagnostic ◽  
Molecular Diagnostic Techniques ◽  
Peste Des Petits Ruminants ◽  
Rt Pcr ◽  
Agar Gel

Peste des petits ruminants (PPR) is a highly contagious and economically important, viral disease of small ruminants caused by the peste des petits ruminants virus (PPRV), which belongs to the genus Morbilivirus in the family Paramyxoviridae. PPR control is achieved mostly through vaccination and/or slaughter of susceptible animals coupled with clinical or laboratory-based diagnosis. Since clinical signs of PPR are not disease-specific and clinical diagnostics is not reliable, it should be confirmed by laboratory testing. Laboratory confirmation of clinical suspicions is made by detection of PPRV in blood, swabs or post-mortem tissues through classical virus isolation (VI), agar gel immunodiffusion (AGID)/agar gel precipitation test (AGPT), counter-immunoelectrophoresis (CIE), immunoperoxidase test (IPT) or enzyme-linked immunosorbent (ELISA) assays. However, these conventional methods have been superseded by more rapid, sensitive and accurate molecular diagnostic techniques based on the amplification of parts of either nucleocapsid (N) or fusion (F) protein gene, such as RT-PCR, real-time RT-PCR, reverse transcription loop-mediated isothermal amplification (RT-LAMP), reverse transcription recombinase polymerase amplification (RT-RPA) and Oxford nanopore MinION technology. Although these molecular diagnostic assays are accurate, rapid and sensitive, they have to be performed in laboratory settings, and samples must be transported under appropriate conditions from the field to the laboratory, which can delay the confirmation of PPRV infection. The recently developed immunochromatographic lateral flow device (IC-LFD) assay can be used in the field (“pen-side”) without the need for expensive equipment, so a well-established laboratory is not required. The control and eventual eradication of PPR is now one of the top priorities for the Food and Agriculture Organization (FAO) and the World Organization for Animal Health (OIE). In 2015, the international community agreed on a global strategy for PPR eradication, setting 2030 as a target date for elimination of the disease

scholarly journals A Recent Update on Advanced Molecular Diagnostic Techniques for COVID-19 Pandemic: An Overview

2021 ◽  
Vol 12 ◽  
Author(s):  
Akanksha Roberts ◽  
Raghuraj Singh Chouhan ◽  
Deepshikha Shahdeo ◽  
Narlawar Sagar Shrikrishna ◽  
Veerbhan Kesarwani ◽  
...  
Keyword(s):  
Reverse Transcription ◽  
Electrochemical Sensors ◽  
Cost Effective ◽  
Surface Enhanced Raman Spectroscopy ◽  
Diagnostic Techniques ◽  
Diagnostic Methods ◽  
Molecular Diagnostic ◽  
Molecular Diagnostic Techniques ◽  
Rt Pcr ◽  
Ongoing Research

Coronavirus disease 2019 (COVID-19), which started out as an outbreak of pneumonia, has now turned into a pandemic due to its rapid transmission. Besides developing a vaccine, rapid, accurate, and cost-effective diagnosis is essential for monitoring and combating the spread of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and its related variants on time with precision and accuracy. Currently, the gold standard for detection of SARS-CoV-2 is Reverse Transcription Polymerase Chain Reaction (RT-PCR), but it lacks accuracy, is time-consuming and cumbersome, and fails to detect multi-variant forms of the virus. Herein, we have summarized conventional diagnostic methods such as Chest-CT (Computed Tomography), RT-PCR, Loop Mediated Isothermal Amplification (LAMP), Reverse Transcription-LAMP (RT-LAMP), as well new modern diagnostics such as CRISPR–Cas-based assays, Surface Enhanced Raman Spectroscopy (SERS), Lateral Flow Assays (LFA), Graphene-Field Effect Transistor (GraFET), electrochemical sensors, immunosensors, antisense oligonucleotides (ASOs)-based assays, and microarrays for SARS-CoV-2 detection. This review will also provide an insight into an ongoing research and the possibility of developing more economical tools to tackle the COVID-19 pandemic.

scholarly journals Camelids and Cattle Are Dead-End Hosts for Peste-des-Petits-Ruminants Virus

Viruses ◽  
2019 ◽  
Vol 11 (12) ◽  
pp. 1133 ◽  
Author(s):  
Claudia Schulz ◽  
Christine Fast ◽  
Ulrich Wernery ◽  
Jörg Kinne ◽  
Sunitha Joseph ◽  
...  
Keyword(s):  
Small Ruminant ◽  
Clinical Signs ◽  
Small Ruminants ◽  
Tissue Tropism ◽  
Peste Des Petits Ruminants ◽  
Dromedary Camels ◽  
Dead End ◽  
Recent Transmission ◽  
Lineage Iv

Peste-des-petits-ruminants virus (PPRV) causes a severe respiratory disease in small ruminants. The possible impact of different atypical host species in the spread and planed worldwide eradication of PPRV remains to be clarified. Recent transmission trials with the virulent PPRV lineage IV (LIV)-strain Kurdistan/2011 revealed that pigs and wild boar are possible sources of PPRV-infection. We therefore investigated the role of cattle, llamas, alpacas, and dromedary camels in transmission trials using the Kurdistan/2011 strain for intranasal infection and integrated a literature review for a proper evaluation of their host traits and role in PPRV-transmission. Cattle and camelids developed no clinical signs, no viremia, shed no or only low PPRV-RNA loads in swab samples and did not transmit any PPRV to the contact animals. The distribution of PPRV-RNA or antigen in lymphoid organs was similar in cattle and camelids although generally lower compared to suids and small ruminants. In the typical small ruminant hosts, the tissue tropism, pathogenesis and disease expression after PPRV-infection is associated with infection of immune and epithelial cells via SLAM and nectin-4 receptors, respectively. We therefore suggest a different pathogenesis in cattle and camelids and both as dead-end hosts for PPRV.

A polymer lab-on-a-chip for reverse transcription (RT)-PCR based point-of-care clinical diagnostics

Lab on a Chip ◽  
2008 ◽  
Vol 8 (12) ◽  
pp. 2121 ◽  
Author(s):  
Soo Hyun Lee ◽  
Sung-Woo Kim ◽  
Ji Yoon Kang ◽  
Chong H. Ahn
Keyword(s):  
Reverse Transcription ◽  
Point Of Care ◽  
Lab On A Chip ◽  
Clinical Diagnostics ◽  
Rt Pcr

scholarly journals Occurrence of peste des petits ruminants and its increasing threat to Europe

2019 ◽  
Vol 75 (02) ◽  
pp. 6209-2019
Author(s):  
WIESŁAW NIEDBALSKI
Keyword(s):  
West Africa ◽  
Indian Subcontinent ◽  
Animal Health ◽  
Small Ruminants ◽  
Global Strategy ◽  
High Morbidity ◽  
Peste Des Petits Ruminants ◽  
The European Union ◽  
Increased Risk ◽  
The World

Peste des petits ruminants (PPR) is a highly contagious disease of small ruminants that leads to high morbidity and mortality, and thereby results in devastating economic consequences to the livestock industry. It is caused by the PPR virus (PPRV), which belongs to the genus Morbilivirus in the family Paramyxoviridae. PPR was first recorded in early 1942 in Ivory Coast, West Africa. In the following years, the disease extended its distribution to other parts of the world and now circulates throughout Northern, Eastern and West Africa, as well as Asia – in particular, China, Central Asia, Eurasia, the Indian subcontinent and the Middle East. The molecular epidemiology of PPRV, based on the sequence comparison of a small region of either the N or the F gene, has revealed the existence of four distinct lineages (I-IV) of the virus. Until June 2018, PPR had never been detected in Europe, with the exception of the European part of Turkish Thrace. However, on 24th June 2018 the Bulgarian authorities reported cases of PPR in sheep in the village of Voden, Bolyarovo municipality of Yambol region, on the border with the Thrace region of Turkey. It was the first occurrence of PPR in Bulgaria and in the European Union (EU). The source of PPR infection in Bulgaria is not clear, it could have been the illegal movement of animals, contaminated materials or humans, e.g. refuges. Due to the increased risk of introduction of PPR from North Africa and Turkey to neighbouring regions, the European Food Safety Authority (EFSA) has recently published a report assessing the risk of PPR spreading in Europe. According to a scientific opinion by the EFSA, the spread of PPRV in the territory of the UE could occur by the illegal transport of infected animals or infected animal products. In order to limit the spread and impact of PPR outbreaks in Europe, rapid detection, movement restriction, prompt culling of infected herds and disinfection measures should be introduced immediately. The control and eventual eradication of PPR is now one of the top priorities for the Food and Agriculture Organization (FAO) and the World Organization for Animal Health (OIE). In 2015, the international community agreed on a global strategy for PPR eradication, setting 2030 as a target date for elimination of the disease.

scholarly journals 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

Animals ◽  
2021 ◽  
Vol 11 (11) ◽  
pp. 3170
Author(s):  
Mana Mahapatra ◽  
Martin Mayora Neto ◽  
Asha Khunti ◽  
Felix Njeumi ◽  
Satya Parida
Keyword(s):  
Genetic Material ◽  
Small Ruminants ◽  
Laboratory Diagnosis ◽  
Effective Control ◽  
Peste Des Petits Ruminants ◽  
Rt Pcr ◽  
Eradication Programme ◽  
Long Distance ◽  
Nucleocapsid Gene ◽  
Field Samples

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.

scholarly journals Rapid Detection of Peste des Petits Ruminants Virus (PPRV) Nucleic Acid Using a Novel Low-Cost Reverse Transcription Loop-Mediated Isothermal Amplification (RT-LAMP) Assay for Future Use in Nascent PPR Eradication Programme

Viruses ◽  
2019 ◽  
Vol 11 (8) ◽  
pp. 699 ◽  
Author(s):  
Mahapatra ◽  
Howson ◽  
Fowler ◽  
Batten ◽  
Flannery ◽  
...  
Keyword(s):  
Nucleic Acid ◽  
Reverse Transcription ◽  
Lamp Assay ◽  
Early Warning Systems ◽  
Isothermal Amplification ◽  
Effective Control ◽  
Peste Des Petits Ruminants ◽  
Rt Pcr ◽  
Eradication Programme ◽  
Loop Mediated Isothermal Amplification

Peste des petits ruminants (PPR) is a disease of small ruminants caused by peste des petits ruminants virus (PPRV), and is endemic in Asia, the Middle East and Africa. Effective control combines the application of early warning systems, accurate laboratory diagnosis and reporting, animal movement restrictions, suitable vaccination and surveillance programs, and the coordination of all these measures by efficient veterinary services. Molecular assays, including conventional reverse transcription-polymerase chain reaction (RT-PCR) and real-time RT-PCR (RT-qPCR) have improved the sensitivity and rapidity of diagnosing PPR. However, currently these assays are only performed within laboratory settings; therefore, the development of field diagnostics for PPR would improve the fast implementation of control policies, particularly when PPR has been targeted to be eradicated by 2030. Loop-mediated isothermal amplification (LAMP) assays are simple to use, rapid, and have sensitivity and specificity within the range of RT-qPCR; and can be performed in the field using disposable consumables and portable equipment. This study describes the development of a novel RT-LAMP assay for the detection of PPRV nucleic acid by targeting the N-protein gene. The RT-LAMP 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. The test displayed 100% concordance with RT-qPCR when considering an RT-qPCR cut-off value of CT >40. Further, the RT-LAMP assay was evaluated using experimental and outbreak samples without prior RNA extraction making it more time and cost-effective. This assay provides a solution for a pen-side, rapid and inexpensive PPR diagnostic for use in the field in nascent PPR eradication programme.

scholarly journals Nucleic Acid Testing of SARS-CoV-2

2021 ◽  
Vol 22 (11) ◽  
pp. 6150
Author(s):  
Hee-Min Yoo ◽  
Il-Hwan Kim ◽  
Seil Kim
Keyword(s):  
Polymerase Chain Reaction ◽  
Nucleic Acid ◽  
Reverse Transcription ◽  
Diagnostic Methods ◽  
Detection Methods ◽  
Molecular Diagnostic ◽  
Nucleic Acid Testing ◽  
Rt Pcr ◽  
Chain Reaction ◽  
Polymerase Chain

The coronavirus disease 2019 (COVID-19) has caused a large global outbreak. It is accordingly important to develop accurate and rapid diagnostic methods. The polymerase chain reaction (PCR)-based method including reverse transcription-polymerase chain reaction (RT-PCR) is the most widely used assay for the detection of SARS-CoV-2 RNA. Along with the RT-PCR method, digital PCR has emerged as a powerful tool to quantify nucleic acid of the virus with high accuracy and sensitivity. Non-PCR based techniques such as reverse transcription loop-mediated isothermal amplification (RT-LAMP) and reverse transcription recombinase polymerase amplification (RT-RPA) are considered to be rapid and simple nucleic acid detection methods and were reviewed in this paper. Non-conventional molecular diagnostic methods including next-generation sequencing (NGS), CRISPR-based assays and nanotechnology are improving the accuracy and sensitivity of COVID-19 diagnosis. In this review, we also focus on standardization of SARS-CoV-2 nucleic acid testing and the activity of the National Metrology Institutes (NMIs) and highlight resources such as reference materials (RM) that provide the values of specified properties. Finally, we summarize the useful resources for convenient COVID-19 molecular diagnostics.

scholarly journals Comparative Diagnostic Performance of a Novel Reverse Transcription Loop-Mediated Isothermal Amplification (RT-LAMP) Kit for the Rapid Detection of SARS-CoV-2

Pathogens ◽  
2021 ◽  
Vol 10 (12) ◽  
pp. 1629
Author(s):  
Alexander Domnich ◽  
Andrea Orsi ◽  
Donatella Panatto ◽  
Vanessa De Pace ◽  
Valentina Ricucci ◽  
...  
Keyword(s):  
Reverse Transcription ◽  
Diagnostic Performance ◽  
Point Of Care ◽  
Lamp Assay ◽  
Laboratory Diagnosis ◽  
Isothermal Amplification ◽  
Molecular Diagnostic ◽  
Rt Pcr ◽  
Laboratory Equipment ◽  
Loop Mediated Isothermal Amplification

Although the reverse transcription-polymerase chain reaction (RT-PCR) is considered a standard-of-care assay for the laboratory diagnosis of SARS-CoV-2, several limitations of this method have been described. Reverse transcription loop-mediated isothermal amplification (RT-LAMP) is an alternative molecular assay and is potentially able to overcome some intrinsic shortcomings of RT-PCR. In this study, we evaluated the diagnostic performance of the novel HG COVID-19 RT-LAMP assay. In this retrospective analysis, a total of 400 routinely collected leftover nasopharyngeal samples with a known RT-PCR result were tested by means of the HG COVID-19 RT-LAMP assay. The overall sensitivity and specificity values of HG COVID-19 RT-LAMP versus RT-PCR were 97.0% (95% CI: 93.6–98.9%) and 98.5% (95% CI: 95.7–99.7%), respectively. Inter-assay agreement was almost perfect (κ = 0.96). Concordance was perfect in samples with high viral loads (cycle threshold < 30). The average time to a positive result on RT-LAMP was 17 min. HG COVID-19 RT-LAMP is a reliable molecular diagnostic kit for detecting SARS-CoV-2, and its performance is comparable to that of RT-PCR. Shorter turnaround times and the possibility of performing molecular diagnostics in the point-of-care setting make it a valuable option for facilities without sophisticated laboratory equipment.

scholarly journals Application of internal control based on recombinant retroviral particles for increasing the PCR assay reliability

2019 ◽  
Vol 64 (4) ◽  
pp. 420-430
Author(s):  
E. G. Fomina ◽  
E. E. Grigorieva ◽  
E. P. Scheslenok ◽  
P. A. Semizhon ◽  
S. V. Tkachev ◽  
...  
Keyword(s):  
Internal Control ◽  
Reverse Transcription ◽  
False Negative ◽  
Molecular Diagnostic ◽  
Egfp Expression ◽  
Probe Design ◽  
Acid Extraction ◽  
Rt Pcr ◽  
Pcr Assay ◽  
Hybridization Probe

Molecular diagnostic tests based on PCR preceded by reverse transcription (RT-PCR) are now used commonly for the detection of viral pathogens with RNA genomes. The application of internal controls to validate the entire process of these assays is necessary to prevent false-negative results caused by inhibition or inefficient extraction. In the present study a strategy to produce a new type of internal control for RT-PCR based on recombinant retroviral particles is described. Cell clones stably producing retroviral particles were established by transfecting GP+env-AM12 packaging cells with constructed MoMuLV-derived retroviral vector pLneo/gfp and subsequent cultivation on selective medium with G418. The egfp gene was used as a target for primers and hybridization probe design for real-time RT-PCR assay and as a marker for flow cytometry analysis of eGFP expression by transfected cells. The developed internal control is stable and ribonuclease resistant, economical to produce, noninfectious and safe for routine use. It closely mimics the natural virus and could be successfully used to monitor all the stages of RT-PCR, including nucleic acid extraction, RNA reverse transcription and amplification.

Sign in / Sign up

Export Citation Format

Share Document