Rapid and visual detection of Lawsonia intracellularis with an improved recombinase polymerase amplification assay combined with a lateral flow dipstick
| Autor: | Kaiyue Tian, Zeng Wang, Jun Zhao, Yuhan Zhang, Huifang Guo, Yanyang Wu, Ning Li |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2019 |
| Předmět: |
Recombinase polymerase amplification
Lateral flow dipstick Swine Lawsonia Bacteria Recombinase Polymerase Amplification Gastrointestinal pathogens Biology Sensitivity and Specificity complex mixtures Lawsonia intracellularis Animals On-site detection Swine Diseases lcsh:Veterinary medicine General Veterinary food and beverages General Medicine Dipstick Amplicon Molecular biology Desulfovibrionaceae Infections Proliferative enteropathy enzymes and coenzymes (carbohydrates) Visual detection Molecular Diagnostic Techniques Nucleic acid lcsh:SF600-1100 Reagent Kits Diagnostic Nucleic Acid Amplification Techniques Research Article |
| Zdroj: | BMC Veterinary Research, Vol 15, Iss 1, Pp 1-10 (2019) BMC Veterinary Research |
| ISSN: | 1746-6148 |
| Popis: | Background Lawsonia intracellularis (L. intracellularis) is the etiologic agent of porcine proliferative enteropathy (PPE), which is reported in many swine breeding countries all over the world, and has caused enormous economic losses in intensive pig production systems. Therefore, the aim of this study was to develop a simple and rapid method for on-site detection of Lawsonia intracellularis (L. intracellularis). As the isothermal recombinase polymerase amplification (RPA) can be performed at a constant temperature and its product is directly observed on a lateral-flow dipstick (LFD) with naked eyes without electrophoresis, the RPA-LFD assay should be useful for field diagnosis of L. intracellularis as well as its detection from clinical samples. Results The established RPA-LFD assay could be finished in 30 min at a wide temperature range of 25 to 40 °C, and the amplicons could be visualized by naked eyes. The developed RPA-LFD assay was specific to dnaA gene of L. intracellularis, and did not detect nucleic acids extracted from other common gastrointestinal pathogens. The minimum detection of this RPA-LFD method was 400 L. intracellularis per reaction, which was as sensitive as conventional PCR. Further, the RPA-LFD assay was performed with 150 clinical fecal samples and the detection results were compared with conventional PCR. Results showed that the coincidence rate of RPA-LFD and conventional PCR was 100%. Conclusions The combined RPA with LFD assay provides a simple, rapid, specific and sensitive alternative for field diagnosis of L. intracellularis infection. |
| Databáze: | OpenAIRE |
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