Development of a Rapid Reverse Transcription-Recombinase Polymerase Amplification Couple Nucleic Acid Lateral Flow Method for Detecting Porcine Epidemic Diarrhoea Virus.
RT-RPA-NALF
RT-qPCR
porcine epidemic diarrhea virus
rapid detection
Journal
Biology
ISSN: 2079-7737
Titre abrégé: Biology (Basel)
Pays: Switzerland
ID NLM: 101587988
Informations de publication
Date de publication:
06 Jul 2022
06 Jul 2022
Historique:
received:
19
05
2022
revised:
30
06
2022
accepted:
05
07
2022
entrez:
14
9
2022
pubmed:
15
9
2022
medline:
15
9
2022
Statut:
epublish
Résumé
Porcine epidemic diarrhea virus (PEDV) infection is an important acute diarrheal disease of swine that results in economic and industrial losses worldwide. The clinical manifestations in infected piglets are severe diarrhea, dehydration with milk curd indigestion, leading to death. The diagnosis of PEDV is essential for monitoring and managing the disease. PEDV can be detected and identified by serology and the nucleic acid of the virus in clinical samples. Therefore, a novel isothermal amplification and detection technique, reverse transcription-recombinase polymerase amplification couple nucleic acid lateral flow (RT-RPA-NALF) was developed for the rapid detection of PEDV. Qualitative reverse transcription-polymerase chain reaction (RT-qPCR) was established as the gold standard assay to compare results. Specific primer pairs and probes were designed, and RT-RPA conditions were optimized to amplify the M gene of PEDV. The established RT-RPA-NALF assay could finish in 25 min at a temperature of 42 °C and the amplicon interpreted by visual detection. The developed RT-RPA-NALF assay was specific to the M gene of PEDV, did not detect other common swine diarrhea pathogens, and showed minimal detection at 10
Identifiants
pubmed: 36101399
pii: biology11071018
doi: 10.3390/biology11071018
pmc: PMC9312133
pii:
doi:
Types de publication
Journal Article
Langues
eng
Subventions
Organisme : Mahidol University (Basic Research Fund: fiscal year 2021)
ID : BRF2-NDFR07/2564
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