scholarly journals Equipment-free recombinase polymerase amplification assay using body heat for visual and rapid point-of-need detection of canine parvovirus 2

2018 ◽  
Vol 39 ◽  
pp. 41-46 ◽  
Author(s):  
Libing Liu ◽  
Jianchang Wang ◽  
Yunyun Geng ◽  
Jinfeng Wang ◽  
Ruiwen Li ◽  
...  
Keyword(s):  
Canine Parvovirus ◽  
Recombinase Polymerase Amplification ◽  
Amplification Assay ◽  
Recombinase Polymerase Amplification Assay ◽  
Body Heat

Development of a recombinase polymerase amplification assay forVibrio parahaemolyticusdetection with an internal amplification control

2018 ◽  
Vol 64 (4) ◽  
pp. 223-230 ◽  
Author(s):  
Huan-Lan Yang ◽  
Shuang Wei ◽  
Ravi Gooneratne ◽  
Anthony N. Mutukumira ◽  
Xue-Jun Ma ◽  
...  
Keyword(s):  
Bacterial Species ◽  
False Negative ◽  
Recombinase Polymerase Amplification ◽  
Internal Amplification Control ◽  
Target Sequence ◽  
Negative Results ◽  
False Negative Results ◽  
Amplification Assay ◽  
Pcr Method ◽  
Recombinase Polymerase Amplification Assay

A novel RPA–IAC assay using recombinase polymerase and an internal amplification control (IAC) for Vibrio parahaemolyticus detection was developed. Specific primers were designed based on the coding sequence for the toxR gene in V. parahaemolyticus. The recombinase polymerase amplification (RPA) reaction was conducted at a constant low temperature of 37 °C for 20 min. Assay specificity was validated by using 63 Vibrio strains and 10 non-Vibrio bacterial species. In addition, a competitive IAC was employed to avoid false-negative results, which co-amplified simultaneously with the target sequence. The sensitivity of the assay was determined as 3 × 103CFU/mL, which is decidedly more sensitive than the established PCR method. This method was then used to test seafood samples that were collected from local markets. Seven out of 53 different raw seafoods were detected as V. parahaemolyticus-positive, which were consistent with those obtained using traditional culturing method and biochemical assay. This novel RPA–IAC assay provides a rapid, specific, sensitive, and more convenient detection method for V. parahaemolyticus.

scholarly journals Evaluation of Recombinase Polymerase Amplification Assay for Detecting Meloidogyne javanica

Plant Disease ◽  
2020 ◽  
Vol 104 (3) ◽  
pp. 801-807
Author(s):  
Yuan-kai Chi ◽  
Wei Zhao ◽  
Meng-di Ye ◽  
Farman Ali ◽  
Tao Wang ◽  
...  
Keyword(s):  
Adult Female ◽  
Marker Gene ◽  
Gene Segment ◽  
Meloidogyne Javanica ◽  
Recombinase Polymerase Amplification ◽  
Lateral Flow Dipstick ◽  
Amplification Assay ◽  
Stage Juvenile ◽  
Sequence Characterized Amplified Regions ◽  
Recombinase Polymerase Amplification Assay

Meloidogyne javanica is one of the most widespread and economically important nematodes in many countries, including China. In this study, a recombinase polymerase amplification (RPA) assay was evaluated for the detection of M. javanica based on the sequences of a sequence-characterized amplified regions marker gene segment. The RPA assay specifically detected M. javanica from individual juvenile or adult female, M. javanica-induced galls, and nematodes in the soil samples. The detection limit of M. javanica RPA assay was 1 pg of purified genomic DNA, 0.01 adult female, or 0.1 second-stage juvenile, which was 10 times more sensitive than conventional PCR assay. Furthermore, combined with lateral flow dipstick (LFD), a visual detection method of LFD-RPA assay was developed, which is suitable for onsite surveys and routine diagnostics. Results indicate that the RPA assay is rapid, sensitive, and reliable for detection and molecular identification of M. javanica.

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