期刊
ANALYST
卷 146, 期 4, 页码 1311-1319出版社
ROYAL SOC CHEMISTRY
DOI: 10.1039/d0an01835j
关键词
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资金
- Fulbright Visiting Scholar Program
- NIH [1R21AI128059, R61 AI140484-01]
Penn-RAMP is a highly sensitive molecular test combining RPA and LAMP techniques, showing superior detection efficiency compared to traditional methods and reducing the likelihood of false negative results.
Sensitive, specific and rapid molecular diagnosis of respiratory diseases in animals and humans is critical to facilitate appropriate control measures and treatment. Conventional polymerase chain reaction (PCR)-based molecular diagnostics requires relatively expensive equipment and trained staff, restricting its use to centralized laboratories with significant delays between sample collection and test results. Herein, we report a highly sensitive, rapid, point-of-need, two-stage-molecular test that requires minimal instrumentation and training. Our test, dubbed Penn-RAMP, combines recombinase polymerase amplification (RPA, 38 degrees C) and loop-mediated isothermal amplification (LAMP, 63 degrees C) in one tube, enabling nested, two-stage isothermal amplification. We demonstrate Penn-RAMP's efficacy by testing for two common viral respiratory diseases of chickens: infectious laryngotracheitis (ILT) and infectious bronchitis (IB) that impose great economic burden worldwide. Test results of clinical samples with our closed-tube Penn-RAMP assays concord with the gold standard quantitative PCR (qPCR) assay; with 10-fold better limit of detection than LAMP and qPCR. Our closed-tube Penn-RAMP assays have the potential to greatly reduce false negatives while requiring minimal instrumentation and training.
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