Peer-reviewed veterinary case report
Rapid detection of duck hepatitis A virus genotype C using reverse transcription loop-mediated isothermal amplification.
- Journal:
- Journal of virological methods
- Year:
- 2014
- Authors:
- Li, Chuanfeng et al.
- Affiliation:
- Shanghai Veterinary Research Institute · China
Plain-English summary
Researchers developed a new test to quickly and accurately detect duck hepatitis A virus genotype C (DHAV-C), which is a virus that can affect ducks. They created a method called reverse transcription loop-mediated isothermal amplification (RT-LAMP) that is much more sensitive than the traditional test, reverse transcriptase-polymerase chain reaction (RT-PCR). In their study, they found that the RT-LAMP test identified positive cases in 26.7% of samples, compared to 20% with RT-PCR, and it can be easily checked for results by looking for color changes. This new test is not only fast and specific but also cost-effective, making it useful for both diagnosing sick ducks and monitoring for the virus in the field. Overall, the RT-LAMP test works well for detecting DHAV-C.
Abstract
A one-step reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay was used and optimized to develop a rapid and sensitive detection system for duck hepatitis A virus genotype C (DHAV-C) RNA. A set of four specific primers was designed against highly conserved sequences located within the 3D gene from DHAV (strain GX1201). Under optimal reaction conditions, the sensitivity of DHAV-C-specific RT-LAMP was 100-fold higher than that of reverse transcriptase-polymerase chain reaction (RT-PCR), with a detection limit of 0.3pg (6.59×10(4) copies) per reaction. No cross-reactivity was observed from the samples of other duck viruses, which is in good accordance with RT-PCR. Furthermore, a positive reaction can be visually inspected by observing turbidity or color change after the addition of SYBR green I dye. The DHAV-C-specific RT-LAMP assay was applied to the samples and compared with RT-PCR. The positive-sample ratios were 26.7% (12 of 45) by RT-LAMP and 20% (9 of 45) by RT-PCR. Therefore, the newly developed RT-LAMP assay is a rapid, specific, sensitive, and cost-effective method of DHAV-C detection. This assay has potential applications in both clinical diagnosis and field surveillance of DHAV-C infection.
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Search related cases →Original publication: https://pubmed.ncbi.nlm.nih.gov/24291148/