Reverse transcription loop-mediated isothermal amplification for the rapid detection of infectious bronchitis virus in infected chicken tissues
| Autor: | Yong Guang Zhang, Jie Zhang, Hao tai Chen, Li qing Ma, Yan ping Ma, Xue peng Cai, Li-na Ma, Yong-sheng Liu, Yao zhong Ding, Xiang tao Liu |
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| Rok vydání: | 2010 |
| Předmět: |
animal structures
viruses Infectious bronchitis virus Molecular Sequence Data Loop-mediated isothermal amplification Polymerase Chain Reaction Sensitivity and Specificity Newcastle disease Article Reverse transcription loop-mediated isothermal amplification (RT-LAMP) Virus Infectious bronchitis virus (IBV) Animals Molecular Biology Reverse Transcription Loop-mediated Isothermal Amplification Poultry Diseases DNA Primers Electrophoresis Agar Gel Base Sequence biology Temperature Outbreak RNA Reverse Transcription Cell Biology biology.organism_classification Virology Reverse transcriptase Detection Organ Specificity embryonic structures Coronavirus Infections Chickens |
| Zdroj: | Molecular and Cellular Probes |
| ISSN: | 0890-8508 |
| DOI: | 10.1016/j.mcp.2009.10.001 |
| Popis: | A reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay targeting the nucleocapsid phosphoprotein gene of infectious bronchitis virus (IBV) was developed. The detection limits for the IBV RT-LAMP assay were 10(1) 50% egg infection dose (EID(50)) per 50 microl of titrated viruses and no cross-reaction of IBV RT-LAMP was found when tested with other viruses including Newcastle disease virus (NDV), avian reovirus (ARV), and infectious laryngotrachietis virus (ILTV) due to their mismatch with IBV RT-LAMP primers. A total of 187 clinical tissues samples (88 blood, 62 kidney and 37 lung) were evaluated and compared to conventional RT-PCR. The sensitivity of RT-LAMP and RT-PCR assays for detecting IBV RNA in clinical specimens was 99.5% and 98.4%, respectively. These findings showed that the RT-LAMP assay has potential usefulness for rapid and sensitive diagnosis in outbreak of IBV. |
| Databáze: | OpenAIRE |
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