scholarly journals Development of a recombinase polymerase amplification assay with lateral flow dipstick for rapid detection of feline parvovirus

2019 ◽  
Vol 271 ◽  
pp. 113679 ◽  
Author(s):  
Zhao-Hua Wang ◽  
Xiao-Jia Wang ◽  
Shao-Hua Hou
Keyword(s):  
Rapid Detection ◽  
Lateral Flow ◽  
Recombinase Polymerase Amplification ◽  
Lateral Flow Dipstick ◽  
Amplification Assay ◽  
Feline Parvovirus ◽  
Recombinase Polymerase Amplification Assay

scholarly journals Development of a Lateral Flow Strip-Based Recombinase Polymerase Amplification Assay for the Detection of Haemonchus contortus in Goat Feces

2021 ◽  
Vol 59 (2) ◽  
pp. 167-171
Author(s):  
Yao-Dong Wu ◽  
Qi-Qi Wang ◽  
Meng Wang ◽  
Hany M. Elsheikha ◽  
Xin Yang ◽  
...  
Keyword(s):  
Detection Limit ◽  
Haemonchus Contortus ◽  
Rapid Detection ◽  
Small Ruminants ◽  
Lateral Flow ◽  
Recombinase Polymerase Amplification ◽  
Lateral Flow Strip ◽  
Specific Sequence ◽  
Amplification Assay ◽  
Recombinase Polymerase Amplification Assay

Haemonchosis remains a significant problem in small ruminants. In this study, the assay of recombinase polymerase amplification (RPA) combined with the lateral flow strip (LFS-RPA) was established for the rapid detection of <i>Haemonchus contortus</i> in goat feces. The assay used primers and a probe targeting a specific sequence in the ITS-2 gene. We compared the performance of the LFS-RPA assay to a PCR assay. The LFS-RPA had a detection limit of 10 fg DNA, which was 10 times less compared to the lowest detection limit obtained by PCR. Out of 24 goat fecal samples, LFS-RPA assay detected <i>H. contortus</i> DNA with 95.8% sensitivity, compared to PCR, 79.1% sensitivity. LFS-RPA assay did not detect DNA from other related helminth species and demonstrated an adequate tolerance to inhibitors present in the goat feces. Taken together, our results suggest that LFS-RPA assay had a high diagnostic accuracy for the rapid detection of <i>H. contortus</i> and merits further evaluation.

scholarly journals Rapid Detection of Burkholderia pseudomallei with a Lateral Flow Recombinase Polymerase Amplification Assay

2019 ◽  
Author(s):  
Yao Peng ◽  
Zheng Xiao ◽  
Biao Kan ◽  
Wei Li ◽  
Wen Zhang ◽  
...  
Keyword(s):  
Rapid Detection ◽  
Burkholderia Pseudomallei ◽  
Limit Of Detection ◽  
Point Of Care ◽  
Lateral Flow ◽  
Recombinase Polymerase Amplification ◽  
Spiked Soil ◽  
Lateral Flow Dipstick ◽  
Life Threatening ◽  
Amplification Assay

AbstractMelioidosis is a severe infectious disease caused by gram-negative, facultative intracellular pathogen Burkholderia pseudomallei (B. pseudomallei). Although cases are increasing reported from other parts of the world, it is an illness of tropical and subtropical climates primarily found in southeast Asia and northern Australia. Because of a 40% mortality rate, this life-threatening disease poses a public health risk in endemic area. Early detection of B. pseudomallei infection benefits greatly to implement effective treatment timely, which is vital for prognosis of a melioidosis patient. In this study, a novel isothermal recombinase polymerase amplification combined with lateral flow dipstick (LF-RPA) assay was established for rapid detection of B.pseudomallei. A set of probe and primers targeting orf2 gene of B. pseudomallei were generated and parameters for the LF-RPA assay were optimized. Result can be easy visualized in 30 minutes with the limit of detection (LoD) as low as 20 femtogram (ca. 25.6 copies) of B. pseudomallei genomic DNA. The assay is highly specific as no cross amplification was observed with 35 non-B. pseudomallei pathogens. Isolates (N=19) from patients of Hainan province of China were retrospectively confirmed by the newly developed method. LoD for B. pseudomallei spiked soil and blood samples were 2.1×103 CFU/g and 4.2×103 CFU/ml respectively. Sensitivity of the LF-RPA assay was comparable to TaqMan Real-Time PCR, however, the LF-RPA assay exhibited a better tolerant to inhibitors in blood than the later. Our results showed that the LF-RPA assay is an alternative to existing PCR-based methods for detection of B. pseudomallei with a potentiality of early accurate diagnosis of melioidosis at point of care or in-field use.

Rapid and visual detection of Meloidogyne hapla using recombinase polymerase amplification combined with a lateral flow dipstick (RPA-LFD) assay

Plant Disease ◽  
2020 ◽  
Author(s):  
Zhiqiang Song ◽  
Xiai Yang ◽  
Xiaowei Zhang ◽  
Mingbao Luan ◽  
Bing Guo ◽  
...  
Keyword(s):  
Rapid Detection ◽  
Lateral Flow ◽  
Recombinase Polymerase Amplification ◽  
Economic Losses ◽  
Meloidogyne Hapla ◽  
Root Knot Nematode ◽  
Resource Limited ◽  
Lateral Flow Dipstick ◽  
Wide Range ◽  
And Control

The northern root-knot nematode, Meloidogyne hapla, is a biotrophic parasite that infects many crops and causes severe economic losses worldwide. Rapid and accurate detection of M. hapla is crucial for disease forecasting and control. We developed a recombinase polymerase amplification combined with a lateral flow dipstick (RPA-LFD) assay for rapid detection of M. hapla. The primers and a probe were designed based on the effector gene 16D10 sequence and were highly specific to M. hapla. The RPA reaction was performed at a wide range of temperatures from 25 to 45°C within 5 to 25 min, and the amplicon was visualized directly on the LFD within 5 min. The detection limits of the RPA-LFD assay were 10-3 female and 10-2 J2/0.5 g of soil, which was 10 times more sensitive than the conventional PCR assay. In addition, the RPA-LFD assay can detect M. hapla from infested plant roots and soil samples, and the entire detection process can be completed within 1.5 h. These results indicate that the RPA-LFD assay is a simple, rapid, specific, sensitive, and visual method that can be used for rapid detection of M. hapla in the field and in resource-limited conditions.

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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