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The detection limit of Pseudomonas aeruginosa in real-time PCR and PCR was 1×10 7 CFU/mL, and the detection limit of Pseudomonas aeruginosa in RPA was 1×10 2 CFU/mL.RPA. In fluorescence quantitative PCR, the higher the concentration of Pseudomonas aeruginosa, the shorter the threshold time and the fewer the number of cycles, i.e. the shorter the time for detecting the positive […]
The results from the present study indicated that RPA assay can be used easily in routine indexing of citrus planting material. To the best of our knowledge, this is the first report on development of a rapid and simplified isothermal detection assay for CYMV and can be utilized as an effective technique in quarantine and budwood certification process.
Experiments confirmed a detection limit as low as 300 fg of DNA and 1.5 × 101 CFU in pure cultures. Furthermore, RPA‐LF exhibited no cross‐reactions with pathogens. Evaluation of the method with food samples indicated that the detection limit was substantially improved to 1.5 × 10° CFU for the original bacterial content in 25 g/mL samples after enrichment for 6
The dengue RT-RPA assay can be successfully performed by simply following the provided written instructions. Deviations from the written protocols did not adversely affect the outcome of the assay. These suggest that the RT-RPA assay is indeed a simple, robust and efficient laboratory method for detection of dengue virus.
Clinical evaluation using 162 ovine and hircine serum and nasal swab samples showed that the performance of both the real-time RT-RPA assay and the conventional RT-RPA assay were comparable to that of real-time RT-PCR.
The direct RT-RPA tests constitute robust, accurate, sensitive and quick methods for detection of potyviruses from recalcitrant plant species. The minimal sample preparation requirements and the possibility of storing RPA reagents without cold chain storage, allow Direct RT-RPA to be adopted in minimally equipped laboratories and with potential use in plant clinic laboratories and seed certification facilities worldwide.
Rapid detection of potyviruses from crude plant extracts Read More
Clinical performance of each internally controlled duplex RPA assay was evaluated by testing 64 archived PCR-positive clinical samples. Compared to real-time PCR, all duplex RPA assays demonstrated 100% diagnostic specificity, with diagnostic sensitivities of 100%, 86.3% and 100% for the S. pneumoniae, N. meningitidis and H. influenzae assays, respectively.
All 274 samples were screened using conventional PCR and 21 (7.66%) samples were positive for T. annulata. All the samples that were RPA positive but PCR negative were sequenced, which confirmed the results of RPA. The highest positive rate was found in Chakwal district, followed by Faisalabad and Jhang. This study demonstrates the application of highly sensitive and specific rapid
The sensitivity of the assay was determined as 3×103 CFU/mL, which is decidedly more sensitive than the established PCR method. This method was then used to test seafood samples which were collected from local market. 7 out of 53 different raw seafoods were detected as V.p positive, which were consistent with those obtained using traditional culturing method and biochemical assay.
The RPA-LFD was able to function at 30-45 °C, and at the temperature of 40 °C, it only took 8 min of amplification to reach the test threshold of amplicons. The established method had both a good specificity and a sensitivity of 100 fg DNA per reaction (20 µL). Regarding practical performance, RPA-LFD performed better than real-time PCR.
