Review: a comprehensive summary of a decade development of the recombinase polymerase amplification

Li, Jia; Macdonald, Joanne; Stetten, Felix von

doi:10.1039/c8an01621f

Cited by 459 publications

(390 citation statements)

References 185 publications

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“…The DNA polymerases extend and displace from 3′ends of the primers and cleaved probe to produce the minor amplicons (from the forward and reverse primers) and a displaced strand. The displaced strand combines with the labelled reverse primer, and leads to the production of a dual-labelled amplicon bound with FAM at 5′ end and Biotin at 3'end (the major amplicon) also called FAM-Biotin linking amplicons [20]. Lateral flow dipstick strips are made of permeable membrane.…”

Section: Endpoint Detection By Lateral Flow Dipstick (Lfd) and Sybr Gmentioning

confidence: 99%

Establishment of a recombinase polymerase amplification (RPA) assay for the detection of Brucella spp. Infection

Gumaa

Cao

et al. 2019

Molecular and Cellular Probes

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A B S T R A C TBrucellosis is a worldwide re-emerging zoonosis. It has an economic impact due to abortion and loss of fertility in livestock. In this study, Real-time recombinase polymerase amplification (RT-RPA-BP26) targeting Brucella spp. bp26 gene and Lateral flow dipstick (LFD-RPA-IS711) combined with SYBR-Green recombinase polymerase amplification (RPA) targeting insertion sequence IS711 region of Brucella spp. bp26 gene, was developed to detect Brucella spp. from different sample types in domestic animals. The sensitivity and specificity of the two developed RPAs were compared with real-time PCR, PCR, and Rose Bengal Plate Test (RBPT). The analytical sensitivity and detection limit of Real-time RPA and LFD RPA were four and six copies per reaction respectively. The detection of six colony forming units (CFU) of the bacteria-bearing construct with the target sequence was within 20 min at 40°C for Real-time RPA and 37°C for LFD RPA. The LFD RPA could work at temperatures between 30 and 35°C and could be completed within 10-30 min. No significant differences were observed when comparing the results from Real-time RPA and LFD RPA to Real-time PCR and PCR. Both methods showed no cross reactivity with Chlamydia abortus, Toxoplasma gondii, Salmonella typhimurium, and Escherichia coli. In conclusion, RPA is a useful and convenient field and point of care test for brucellosis.

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Section: Endpoint Detection By Lateral Flow Dipstick (Lfd) and Sybr Gmentioning

confidence: 99%

Establishment of a recombinase polymerase amplification (RPA) assay for the detection of Brucella spp. Infection

Gumaa

Cao

et al. 2019

Molecular and Cellular Probes

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“…Further studies are crucial to improve the assays to be able to distinguish field virulent virus strains from vaccine strains which needed to choose appropriate control measures. Also, a simple and field applicable method to extract pathogens' nucleic acids will greatly facilitate using RPA-NALF assays for on-site diagnosis [34].…”

Section: Discussionmentioning

confidence: 99%

Recombinase polymerase amplification–nucleic acid lateral flow immunoassays for Newcastle disease virus and infectious bronchitis virus detection

et al. 2019

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Newcastle disease virus (NDV) and infectious bronchitis virus (IBV) are two poultry pathogens affecting the respiratory tract of chickens, and cause major economic losses in the industry. Rapid detection of these viruses is crucial to inform implementation of appropriate control measures. The objective of our study is developing a simple, rapid and field applicable recombinase polymerase amplification (RPA)-nucleic acid lateral flow (NALF) immunoassay for detection of NDV and IBV. Isothermal amplification of the matrix protein (M) gene of NDV and the nucleoprotein (N) gene of IBV was implemented via recombinase polymerase amplification at 38 °C for 40 min and 20 min, respectively using modified labeled primers. NALF device used in this study utilizes antibodies for detection of labeled RPA amplicons. The results revealed that RPA-NALF immunoassays can detect both viruses after 40 min at 38 °C and only NDV after 20 min. The limit of detection (LOD) was 10 genomic copies/RPA reaction. The assays results on clinical samples collected from diseased chicken farms demonstrated a good performance in comparison with quantitative real time reverse transcription polymerase chain reaction (qRT-PCR). The assays established in this study can facilitate rapid, on-site molecular diagnosis of suspected cases of ND and IB viral infections as the results can be detected by the naked eye without the need for measuring fluorescence. Furthermore, the NALF device could be adapted to detect other infectious agents.

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“…Several RPA detection methods have been developed so far; in addition to animal virus, plant virus, fungi, bacteria, mycoplasma, and parasite RPA detection methods have been developed in recent years (Cui and Zhao, 2018;Karakkat and Hockemeyer, 2018;Zeng and Luo, 2019;Zhao and Hou, 2018;Hu and Zhong, 2019). Although the RPA technology has not been promoted on the market, some reports have verified its capabilities and its potential to eclipse PCR for further revolutions the life sciences (Li and Macdonald, 2019). With the development of RPA-LFD, it is possible to achieve field detection.…”

Section: Discussionmentioning

confidence: 99%

“…It relies on a synthetically engineered adaptation of a natural cellular process called homologous recombination, comprising three key proteins (recombinase, recombinase loading factor and single-stranded binding protein). The enzymatic activity catalyzes the reaction at a single optimum temperature between 37 ℃ and 42 ℃ (Daher and Stewart, 2016;Li and Macdonald et al, 2019). RPA can routinely generate results within 20 min or less.…”

Section: Introductionmentioning

confidence: 99%

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

Wang

Hou

2019

Journal of Virological Methods

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Feline panleukopenia caused by feline parvovirus (FPV), a single-stranded DNA virus, is typically highly contagious and often presents with lethal syndrome. The broad spectrum of possible hosts suggests its potential for transmission from animal to person through close contact with pets. FPV thus serves as an example of the importance of new rapid point-of-care field diagnostic tools for the control and prevention of transmission, especially among rare wild animals and pet cats. Recombinase polymerase amplification (RPA), as a real-time and isothermal method, could be a more affordable alternative to PCR when combined with a lateral flow dipstick (LFD) indicator. In this study, we report a novel FPV lateral flow dipstick RPA (LFD-RPA) instant detection method capable of detecting a range of different FPV strains. The LFD-RPA assay consists of specific primers, probe, and nucleic acid strip. It is capable of detecting 10 2 copies of target nucleic acid per reaction, which is one order of magnitude higher than the sensitivity of traditional PCR. The most suitable reaction conditions for this assay are at 38 ℃ for 15 min. This paper develops an efficient visual detection system that can eliminate the need for professional staff and expensive and sophisticated equipment for field detection.

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Review: a comprehensive summary of a decade development of the recombinase polymerase amplification

Abstract: RPA is a versatile complement or replacement of PCR, and now is stepping into practice.

Cited by 459 publications

References 185 publications

Establishment of a recombinase polymerase amplification (RPA) assay for the detection of Brucella spp. Infection

Establishment of a recombinase polymerase amplification (RPA) assay for the detection of Brucella spp. Infection

Recombinase polymerase amplification–nucleic acid lateral flow immunoassays for Newcastle disease virus and infectious bronchitis virus detection

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

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